LOCUS       pEN-L4-PAGRO1-SpCas9-T3T-R1_        7126 bp    DNA     circular UNA 22-MAR-2023
DEFINITION  .
ACCESSION   urn.local...i-co498nk
VERSION     urn.local...i-co498nk
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     attL4           1..97
                     /created_by="mamee"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2023.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attL4"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:lr-eztz0t4"">attL4</a>"
                     /label="attL4"
     promoter        104..378
                     /%_Identity=100
                     /Motif="CACGAACCCAGTGGACATAAGCCTGTTCGGTTCGTAAGCTGTAATGCAAG
                     TAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACG
                     GCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGGGGTACAGTCTATGCC
                     TCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGC
                     AGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAAACATC"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="SpR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; P1 (<a
                     href=""urn:local:thjac:2r-c4nnd56"">SpR</a>)"
                     /modified_by="mamee"
                     /label="pAgro1"
     CDS             413..4519
                     /created_by="lapau"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="SpCas9"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:lapau:13-ciehhao"">SpCas9</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:3/-3
                     (adjusted to 99.85%; Adjusted-Interval-0:0/0); <a
                     href=""urn:local:warddecaestecker:2q-c8811q6"">SpCas9</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:3/-3
                     (adjusted to 99.85%; Adjusted-Interval-0:0/0); zCAS9 (<a
                     href=""urn:local:thjac:i-cj7qjpf"">ZmCas9
                     translation</a>)"
                     /Alternative_Match="99.93%; Adjusted-Interval-0:3/-3
                     (adjusted to 99.78%; Adjusted-Interval-0:0/0); <a
                     href=""urn:local:warddecaestecker:2p-c8811q6"">SpCas9(D10A
                     )</a>"
                     /Alternative_Match="99.93%; Adjusted-Interval-0:3/-3
                     (adjusted to 99.78%; Adjusted-Interval-0:0/0); <a
                     href=""urn:local:warddecaestecker:2o-c8811q6"">SpCas9(H840
                     A)</a>"
                     /Alternative_Match="99.85%; Adjusted-Interval-0:3/-3
                     (adjusted to 99.71%; Adjusted-Interval-0:0/0); <a
                     href=""urn:local:warddecaestecker:2s-c8811q6"">SpCas9(D10A
                     ,H840A)</a>"
                     /label="SpCas9"
     terminator      4524..4575
                     /created_by="lapau"
                     /label="T3 terminator"
     attR1           4581..4737
                     /created_by="mamee"
                     /Color="#00cccc"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2023.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="attR1"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:y-f0o11e8"">attR1</a>"
                     /label="attR1"
     primer          complement(4797..4817)
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pCDB-Cas9 - GWM13R"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13R
                     (pCDB-Cas9 - GWM13R)"
                     /label="GWM13R"
     promoter        4878..4928
                     /created_by="chgai"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="pCG034_pKan_Kan-CBE"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ll-ch8iv1b"">pCG034_pKan_Kan-CBE</a
                     >)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lo-ch8iv1b"">pCG040_ABE-Nuc-NLS_Kan
                     -reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lr-ch8iv1b"">pCG043_Kan-ABE-Dest_co
                     ntrol_ccdB</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lu-ch8iv1c"">pCG046_ABE-LbCas12a-NL
                     S_Kan_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:kx-ch8iv19"">pCG005_pGGD-pKan-KanR-
                     E</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ky-ch8iv19"">pCG006_pGGA-Kan_promot
                     er-B</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lm-ch8iv1b"">pCG035_pKan_Kan</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:ln-ch8iv1b"">pCG039_ABE-N7-NLS_Kan-
                     reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lp-ch8iv1b"">pCG041_ABE-SV40-NLS_Ka
                     n-reporter_control</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Kan_promoter (<a
                     href=""urn:local:chgai:lw-ch8iv1c"">pCG054_Kan_CBE_control
                     </a>)"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2023.0.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (<a
                     href=""urn:local:.:8j-dgycryy"">KanR</a>)"
                     /label="kan"
     misc_feature    4929..5738
                     /modified_by="wacae"
                     /note="Derived using Geneious Prime 2020.2.4 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="KanR"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:thjac:2o-c4nnd56"">KanR</a>"
                     /label="KanR"
                     /note="Geneious type: Bacterial Selection"
     rep_origin      5885..6473
                     /created_by="thjac"
                     /modified_by="mamee"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of  replication"
                     /Transferred_Similarity="99.49%"
                     /Primary_Match="99.49%; Adjusted-Interval-0:0/0;
                     ColE1/pMB1/pBR322/pUC Ori (<a
                     href=""urn:local:.:8d-dk4mzop"">high-copy-number
                     ColE1/pMB1/pBR322/pUC origin of  replication</a>)"
                     /label="ColE1/pMB1/pBR322/pUC Ori"
     primer          7072..7092
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pCDB-Cas9 - GWM13F"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; GWM13F
                     (pCDB-Cas9 - GWM13F)"
                     /label="GWM13F"
ORIGIN      
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgataagcaa
       61 tgctttttta taatgccaac tttgtataga aaagttgcta cctcacgaac ccagtggaca
      121 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
      181 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
      241 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
      301 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
      361 aaaacaaagt taaacatcaa cagtattcag tcgactggta ccaacaggct ccatggataa
      421 gaaatactca ataggcttag atatcggcac aaatagcgtc ggatgggcgg tgatcactga
      481 tgaatataag gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat
      541 caaaaaaaat cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg
      601 tctcaaacgg acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca
      661 ggagattttt tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga
      721 gtcttttttg gtggaagaag acaagaagca tgaacgtcat cctatttttg gaaatatagt
      781 agatgaagtt gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt
      841 agattctact gataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa
      901 gtttcgtggt cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa
      961 actatttatc cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc
     1021 aagtggagta gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga
     1081 aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc
     1141 tttgtcattg ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa
     1201 attacagctt tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg
     1261 agatcaatat gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc
     1321 agatatccta agagtaaata ctgaaataac taaggctccc ctatcagctt caatgattaa
     1381 acgctacgat gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact
     1441 tccagaaaag tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat
     1501 tgatggggga gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat
     1561 ggatggtact gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg
     1621 gacctttgac aacggctcta ttccccatca aattcacttg ggtgagctgc atgctatttt
     1681 gagaagacaa gaagactttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat
     1741 cttgactttt cgaattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc
     1801 atggatgact cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga
     1861 taaaggtgct tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc
     1921 aaatgaaaaa gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga
     1981 attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga
     2041 acagaagaaa gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca
     2101 attaaaagaa gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt
     2161 tgaagataga tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga
     2221 taaagatttt ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt
     2281 gaccttattt gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt
     2341 tgatgataag gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc
     2401 tcgaaaattg attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt
     2461 gaaatcagat ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac
     2521 atttaaagaa gacattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca
     2581 tattgcaaat ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt
     2641 tgttgatgaa ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat
     2701 ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg
     2761 aatcgaagaa ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa
     2821 tactcaattg caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta
     2881 tgtggaccaa gaattagata ttaatcgttt aagtgattat gatgtcgatc acattgttcc
     2941 acaaagtttc cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa
     3001 tcgtggtaaa tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg
     3061 gagacaactt ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc
     3121 tgaacgtgga ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga
     3181 aactcgccaa atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata
     3241 cgatgaaaat gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt
     3301 ttctgacttc cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca
     3361 tgcccatgat gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa
     3421 acttgaatcg gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc
     3481 taagtctgag caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat
     3541 gaacttcttc aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat
     3601 cgaaactaat ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt
     3661 gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg
     3721 cggattctcc aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa
     3781 aaaagactgg gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt
     3841 cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt
     3901 actagggatc acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga
     3961 agctaaagga tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct
     4021 ttttgagtta gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg
     4081 aaatgagctg gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga
     4141 aaagttgaag ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa
     4201 gcattattta gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc
     4261 agatgccaat ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg
     4321 tgaacaagca gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc
     4381 ttttaaatat tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt
     4441 agatgccact cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag
     4501 tcagctagga ggtgactgat cagaataagc aaaccccttg ggttccctct ttaggagtct
     4561 gaggggtttt ttgcagtatg caagtttgta caaaaaagtt gaacgagaaa cgtaaaatga
     4621 tataaatatc aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa
     4681 aacacaacat atgcagtcac tatgaatcaa ctacttagat ggtattagtg acctgtagaa
     4741 ttcgagctct agagctgcag ggcggccgcg atatccccta tagtgagtcg tattacatgg
     4801 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     4861 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     4921 ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg
     4981 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     5041 tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5101 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     5161 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     5221 cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     5281 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     5341 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     5401 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     5461 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     5521 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     5581 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     5641 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     5701 aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt
     5761 aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc
     5821 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5881 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5941 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6001 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6061 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6121 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6181 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6241 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     6301 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6361 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6421 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6481 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6541 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6601 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6661 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6721 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     6781 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     6841 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     6901 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     6961 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     7021 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
     7081 cggccagtct taagctcggg cccgcgttaa cgctaccatg gagctc
//
