LOCUS pCDB-Cas9 8957 bp DNA circular UNA 23-FEB-2023
DEFINITION .
ACCESSION urn.local...b-cixsr3m
VERSION urn.local...b-cixsr3m
KEYWORDS .
SOURCE
ORGANISM .
FEATURES Location/Qualifiers
attL1 1..100
/Color="#00cccc"
/created_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="attL1"
/Transferred_Similarity="99.00%"
/Primary_Match="99.00%; Adjusted-Interval-0:0/0; attL1"
/label="attL1"
promoter 107..970
/regulatory_class="promoter"
/gene="CaMV 35S"
/note="internal BsaI recognition site removed"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="pCaMV 35S - Golden Gate"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; p35S (pCaMV 35S - Golden
Gate)"
/label="p35S"
Linker 975..998
/gene="B-dummy"
/note="default module for GreenGate"
/modified_by="mamee"
/label="B-dummy 5'UTR"
CDS 1005..5108
/modified_by="mamee"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="AtCas9"
/Transferred_Similarity="100.00%"
/Adjusted_Transferred_Similarity="99.13%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/-36
(adjusted to 99.13%; Adjusted-Interval-0:0/0); AtCas9"
/label="AtCAS9"
NLS 5121..5141
/created_by="warddecaestecker"
/modified_by="wacae"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on protein similarity"
/Transferred_From="NLS SV40"
/Transferred_Translation_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; SV40 (NLS SV40)"
/label="SV40"
Linker 5149..5194
/gene="D-dummy"
/note="default module for GreenGate"
/modified_by="mamee"
/label="D-dummy 3'UTR"
terminator 5199..5845
/regulatory_class="terminator"
/note="RBCS terminator (from Pisum sativum)"
/modified_by="mamee"
/label="tRbcS"
promoter 5852..6239
/vntifkey=29
/modified_by="thjac"
/modified_by="wacae"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="AtU6-26"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; AtU6-26"
/label="AtU6-26"
Scaffold 6258..6340
/%_Identity=100
/Motif="GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC
TTGAAAAAGTGGCACCGAGTCGGTGCTTTTTTT"
/annotation_group="Scaffold: 451 -> 533"
/modified_by="wacae"
/note="Derived using Geneious Prime 2020.2.2 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="SpCas9 Scaffold"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; Scaffold
(SpCas9 Scaffold)"
/label="Scaffold"
attL2 complement(6346..6445)
/created_by="mamee"
/Color="#00cccc"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="attL2"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; attL2"
/label="attL2"
primer complement(6483..6503)
/label="GWM13R"
promoter 6564..6614
/created_by="chgai"
/X-Transferred_From="pCG034_pKan_Kan-CBE"
/X-Transferred_Similarity="100.00%"
/X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG034_pKan_Kan-CBE)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG040_ABE-Nuc-NLS_Kan
-reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG043_Kan-ABE-Dest_co
ntrol_ccdB)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG046_ABE-LbCas12a-NL
S_Kan_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG005_pGGD-pKan-KanR-
E)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG006_pGGA-Kan_promot
er-B)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG035_pKan_Kan)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG039_ABE-N7-NLS_Kan-
reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG041_ABE-SV40-NLS_Ka
n-reporter_control)"
/X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
Kan_promoter (pCG054_Kan_CBE_control
)"
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="KanR"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; kan (KanR)"
/label="kan"
misc_feature 6615..7424
/modified_by="mamee"
/note="Derived using Geneious Prime 2022.1.1 'Annotate
from Database' based on nucleotide similarity"
/Transferred_From="KanR"
/Transferred_Similarity="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0; KanR"
/label="KanR"
/note="Geneious type: Bacterial Selection"
rep_origin 7571..8159
/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="100.00%"
/Primary_Match="100.00%; Adjusted-Interval-0:0/0;
ColE1/pMB1/pBR322/pUC Ori (high-copy-number
ColE1/pMB1/pBR322/pUC origin of replication)"
/label="ColE1/pMB1/pBR322/pUC Ori"
misc_feature 8708..8875
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
primer 8926..8946
/label="GWM13F"
ORIGIN
1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
61 tgctttttta taatgccaac tttgtacaaa aaagcaggct taacctccga tccattcggt
121 ccccagatta gccttttcaa tttcagaaag aatgctaacc cacagatggt tagagaggct
181 tacgcagcac gtctcatcaa gacgatctac ccgagcaata atctccagga aatcaaatac
241 cttcccaaga aggttaaaga tgcagtcaaa agattcagga ctaactgcat caagaacaca
301 gagaaagata tatttctcaa gatcagaagt actattccag tatggacgat tcaaggcttg
361 cttcacaaac caaggcaagt aatagagatt ggagtctcta aaaaggtagt tcccactgaa
421 tcaaaggcca tggagtcaaa gattcaaata gaggacctaa cagaactcgc cgtaaagact
481 ggcgaacagt tcatacagag tctcttacga ctcaatgaca agaagaaaat cttcgtcaac
541 atggtggagc acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac
601 caaagggcaa ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat
661 tgcccagcta tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa
721 tgccatcatt gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc
781 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
841 tcaaagcaag tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac
901 tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga gagaacacgg
961 gggacgagct aacagtattc agtcgactgg taccaacagg ctccatggat aagaagtact
1021 ctatcggact cgatatcgga actaactctg tgggatgggc tgtgatcacc gatgagtaca
1081 aggtgccatc taagaagttc aaggttctcg gaaacaccga taggcactct atcaagaaaa
1141 accttatcgg tgctctcctc ttcgattctg gtgaaactgc tgaggctacc agactcaaga
1201 gaaccgctag aagaaggtac accagaagaa agaacaggat ctgctacctc caagagatct
1261 tctctaacga gatggctaaa gtggatgatt cattcttcca caggctcgaa gagtcattcc
1321 tcgtggaaga agataagaag cacgagaggc accctatctt cggaaacatc gttgatgagg
1381 tggcatacca cgagaagtac cctactatct accacctcag aaagaagctc gttgattcta
1441 ctgataaggc tgatctcagg ctcatctacc tcgctctcgc tcacatgatc aagttcagag
1501 gacacttcct catcgagggt gatctcaacc ctgataactc tgatgtggat aagttgttca
1561 tccagctcgt gcagacctac aaccagcttt tcgaagagaa ccctatcaac gcttcaggtg
1621 tggatgctaa ggctatcctc tctgctaggc tctctaagtc aagaaggctt gagaacctca
1681 ttgctcagct ccctggtgag aagaagaacg gacttttcgg aaacttgatc gctctctctc
1741 tcggactcac ccctaacttc aagtctaact tcgatctcgc tgaggatgca aagctccagc
1801 tctcaaagga tacctacgat gatgatctcg ataacctcct cgctcagatc ggagatcagt
1861 acgctgattt gttcctcgct gctaagaacc tctctgatgc tatcctcctc agtgatatcc
1921 tcagagtgaa caccgagatc accaaggctc cactctcagc ttctatgatc aagagatacg
1981 atgagcacca ccaggatctc acacttctca aggctcttgt tagacagcag ctcccagaga
2041 agtacaaaga gattttcttc gatcagtcta agaacggata cgctggttac atcgatggtg
2101 gtgcatctca agaagagttc tacaagttca tcaagcctat cctcgagaag atggatggaa
2161 ccgaggaact cctcgtgaag ctcaatagag aggatcttct cagaaagcag aggaccttcg
2221 ataacggatc tatccctcat cagatccacc tcggagagtt gcacgctatc cttagaaggc
2281 aagaggattt ctacccattc ctcaaggata acagggaaaa gattgagaag attctcacct
2341 tcagaatccc ttactacgtg ggacctctcg ctagaggaaa ctcaagattc gcttggatga
2401 ccagaaagtc tgaggaaacc atcacccctt ggaacttcga agaggtggtg gataagggtg
2461 ctagtgctca gtctttcatc gagaggatga ccaacttcga taagaacctt ccaaacgaga
2521 aggtgctccc taagcactct ttgctctacg agtacttcac cgtgtacaac gagttgacca
2581 aggttaagta cgtgaccgag ggaatgagga agcctgcttt tttgtcaggt gagcaaaaga
2641 aggctatcgt tgatctcttg ttcaagacca acagaaaggt gaccgtgaag cagctcaaag
2701 aggattactt caagaaaatc gagtgcttcg attcagttga gatttctggt gttgaggata
2761 ggttcaacgc atctctcgga acctaccacg atctcctcaa gatcattaag gataaggatt
2821 tcttggataa cgaggaaaac gaggatatct tggaggatat cgttcttacc ctcaccctct
2881 ttgaagatag agagatgatt gaagaaaggc tcaagaccta cgctcatctc ttcgatgata
2941 aggtgatgaa gcagttgaag agaagaagat acactggttg gggaaggctc tcaagaaagc
3001 tcattaacgg aatcagggat aagcagtctg gaaagacaat ccttgatttc ctcaagtctg
3061 atggattcgc taacagaaac ttcatgcagc tcatccacga tgattctctc acctttaaag
3121 aggatatcca gaaggctcag gtttcaggac agggtgatag tctccatgag catatcgcta
3181 acctcgctgg atctcctgca atcaagaagg gaatcctcca gactgtgaag gttgtggatg
3241 agttggtgaa ggtgatggga aggcataagc ctgagaacat cgtgatcgaa atggctagag
3301 agaaccagac cactcagaag ggacagaaga actctaggga aaggatgaag aggatcgagg
3361 aaggtatcaa agagcttgga tctcagatcc tcaaagagca ccctgttgag aacactcagc
3421 tccagaatga gaagctctac ctctactacc tccagaacgg aagggatatg tatgtggatc
3481 aagagttgga tatcaacagg ctctctgatt acgatgttga tcatatcgtg ccacagtcat
3541 tcttgaagga tgattctatc gataacaagg tgctcaccag gtctgataag aacaggggta
3601 agagtgataa cgtgccaagt gaagaggttg tgaagaaaat gaagaactat tggaggcagc
3661 tcctcaacgc taagctcatc actcagagaa agttcgataa cttgactaag gctgagaggg
3721 gaggactctc tgaattggat aaggcaggat tcatcaagag gcagcttgtg gaaaccaggc
3781 agatcactaa gcacgttgca cagatcctcg attctaggat gaacaccaag tacgatgaga
3841 acgataagtt gatcagggaa gtgaaggtta tcaccctcaa gtcaaagctc gtgtctgatt
3901 tcagaaagga tttccaattc tacaaggtga gggaaatcaa caactaccac cacgctcacg
3961 atgcttacct taacgctgtt gttggaaccg ctctcatcaa gaagtatcct aagctcgagt
4021 cagagttcgt gtacggtgat tacaaggtgt acgatgtgag gaagatgatc gctaagtctg
4081 agcaagagat cggaaaggct accgctaagt atttcttcta ctctaacatc atgaatttct
4141 tcaagaccga gattaccctc gctaacggtg agatcagaaa gaggccactc atcgagacaa
4201 acggtgaaac aggtgagatc gtgtgggata agggaaggga tttcgctacc gttagaaagg
4261 tgctctctat gccacaggtg aacatcgtta agaaaaccga ggtgcagacc ggtggattct
4321 ctaaagagtc tatcctccct aagaggaact ctgataagct cattgctagg aagaaggatt
4381 gggaccctaa gaaatacggt ggtttcgatt ctcctaccgt ggcttactct gttctcgttg
4441 tggctaaggt tgagaaggga aagagtaaga agctcaagtc tgttaaggaa cttctcggaa
4501 tcactatcat ggaaaggtca tctttcgaga agaacccaat cgatttcctc gaggctaagg
4561 gatacaaaga ggttaagaag gatctcatca tcaagctccc aaagtactca ctcttcgaac
4621 tcgagaacgg tagaaagagg atgctcgctt ctgctggtga gcttcaaaag ggaaacgagc
4681 ttgctctccc atctaagtac gttaactttc tttacctcgc ttctcactac gagaagttga
4741 agggatctcc agaagataac gagcagaagc aacttttcgt tgagcagcac aagcactact
4801 tggatgagat catcgagcag atctctgagt tctctaaaag ggtgatcctc gctgatgcaa
4861 acctcgataa ggtgttgtct gcttacaaca agcacagaga taagcctatc agggaacagg
4921 cagagaacat catccatctc ttcaccctta ccaacctcgg tgctcctgct gctttcaagt
4981 acttcgatac aaccatcgat aggaagagat acacctctac caaagaagtg ctcgatgcta
5041 ccctcatcca tcagtctatc actggactct acgagactag gatcgatctc tcacagctcg
5101 gtggtgattc aagggctgat cctaagaaga agaggaaggt ttgatcaggt ggatcctaga
5161 taacctttac cttcattccc tgtaaattgg taccctgcaa agctttcgtt cgtatcatcg
5221 gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca ccagaatcct
5281 actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt tgttgtgctt
5341 gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg
5401 agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat tatttgtttt
5461 ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac attttgtttt
5521 gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag gagtaaaaca
5581 cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca gacctagaaa
5641 agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt tatgaacttt
5701 cctttatgta attttccaga atccttgtca gattctaatc attgctttat aattatagtt
5761 atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta tgacttgcca
5821 attgattgac aacatgcatc aatcgactac tctttttttc ttcttcttcg ttcatacagt
5881 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa
5941 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag
6001 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata
6061 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
6121 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
6181 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattgg
6241 gagacccgag ggtctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
6301 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt gtattaccca gctttcttgt
6361 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
6421 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
6481 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
6541 ttgcacaaga taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta
6601 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
6661 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
6721 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
6781 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
6841 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
6901 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
6961 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
7021 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
7081 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
7141 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
7201 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
7261 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
7321 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
7381 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
7441 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
7501 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
7561 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
7621 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
7681 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
7741 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
7801 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
7861 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
7921 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
7981 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
8041 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
8101 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
8161 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
8221 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
8281 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
8341 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
8401 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
8461 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
8521 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
8581 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
8641 acaacagata aaacgaaagg cccagacttc cgactgagcc tttcgtttta tttgatgcct
8701 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
8761 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
8821 acgaaaggcc cagacttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
8881 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
8941 gtcttaagct cgggccc
//