LOCUS pET28-CBE-nC9s 10713 bp DNA circular SYN 03-JUL-2025 DEFINITION pET28-CBE-nC9s (G803): Ligation of Nhe-Xho* into F611pet28-CBE-nC9* ACCESSION pET28-CBE-nC9s KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct COMMENT DNA Cloning Service, https://dna-cloning.com/vectors/puc/pET28-CBE-nC9s FEATURES Location/Qualifiers CDS 20..37 /label="T7 promoter" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 36..66 /label="lac\operator" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 119..136 /label="His-Tag" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" misc_feature 136 /label="'TEV" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 137..707 /label="GAM" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 708..1391 /label="APOBEC1" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 1391..1481 /label="SGGS Linker'" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 1487..5590 /label="nCas9" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" misc_feature 1511..1513 /label="D10A" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 5597..5614 /label="His-Tag" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 5615..5617 /label="Stop" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS 5765..6220 /label="f1 ori" /note="color: #8B8F86" /ApEinfo_fwdcolor="#8B8F86" /ApEinfo_revcolor="#8B8F86" CDS complement(6315..7127) /label="kan\sequence" /note="color: #3557B7" /ApEinfo_fwdcolor="#3557B7" /ApEinfo_revcolor="#3557B7" CDS 7249..7837 /label="ori" /note="color: #8B8F86" /ApEinfo_fwdcolor="#8B8F86" /ApEinfo_revcolor="#8B8F86" CDS 8023..8162 /label="bom" /note="color: #8B8F86" /ApEinfo_fwdcolor="#8B8F86" /ApEinfo_revcolor="#8B8F86" CDS 8264..8455 /label="rop" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" CDS 9230..9251 /label="CAP binding site" /note="color: #7B5EA7" /ApEinfo_fwdcolor="#7B5EA7" /ApEinfo_revcolor="#7B5EA7" CDS complement(9267..10346) /label="lac\I" /note="color: #D9A400" /ApEinfo_fwdcolor="#D9A400" /ApEinfo_revcolor="#D9A400" ORIGIN 1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 61 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca 121 tcatcatcat catcacatgg ctaaaccggc aaaacgtatc aagtctgcgg ctgcagctta 181 cgttccgcag aaccgcgacg ccgtaatcac cgatatcaaa cgtattggtg acctgcagcg 241 tgaagcgtcc cgtctggaaa ccgagatgaa cgacgctatc gcagaaatca ctgagaaatt 301 cgctgcgcgc atcgctccga tcaaaaccga tatcgaaact ctgagcaagg gcgttcaggg 361 ttggtgcgaa gcaaaccgtg acgaactgac caacggcggt aaagtgaaaa ctgctaacct 421 ggttaccggc gacgtatctt ggcgtgttcg tccaccgtct gtctccatcc gcggtatgga 481 cgcagttatg gaaacactgg aacgtctggg tttacaacgt tttattcgta ctaaacagga 541 aatcaacaaa gaagctatcc tgctggagcc gaaggcggta gctggcgttg ctggtatcac 601 cgtgaaatcc ggcatcgaag atttcagcat cattccgttc gaacaggaag ccggtatctc 661 tggctcagaa actccgggta cctctgagtc cgcaactccg gaatcttcca gcgaaaccgg 721 ccctgttgcg gtagacccga cgctgcgccg tcgtatcgaa ccgcacgaat tcgaggtatt 781 ctttgacccg cgtgaactgc gcaaagaaac ttgtctgttg tacgaaatca actggggtgg 841 tcgtcactct atctggcgtc ataccagtca gaatactaac aaacatgttg aggttaactt 901 catcgagaag ttcaccactg aacgctactt ttgcccgaac acccgttgct ccatcacttg 961 gttcctgtct tggagcccat gtggcgaatg ctctcgtgct attaccgaat tcctgtcccg 1021 ttatccgcac gttactctgt tcatctacat cgcacgcctg taccaccacg ctgatccgcg 1081 taaccgtcag ggtctgcgtg accttatctc ttccggcgtg accatccaga tcatgactga 1141 acaggagtct ggttactgct ggcgcaactt tgttaactac agcccgtcca acgaagcgca 1201 ctggccgcgt tacccgcatc tgtgggtacg tctgtatgtt ctggaactgt actgcatcat 1261 tctgggcctg ccgccatgtc tcaacatcct gcgtcgcaaa caaccgcagc tgaccttctt 1321 cactatcgca ctgcagtctt gccactacca gcgtctgccg ccgcacatcc tgtgggctac 1381 tggtctgaaa tccggcggaa gcagtggcgg gtccagcggc tcagaaactc cagggacgtc 1441 cgagagcgcc actcccgagt cgtccggcgg ttctagcgga ggatccgata agaaatactc 1501 aataggctta gccatcggca caaatagcgt cggatgggcg gtgatcactg atgaatataa 1561 ggttccgtct aaaaagttca aggttctggg aaatacagac cgccacagta tcaaaaaaaa 1621 tcttataggg gctcttttat ttgacagtgg agagacagcg gaagcgactc gtctcaaacg 1681 gacagctcgt agaaggtata cacgtcggaa gaatcgtatt tgttatctac aggagatttt 1741 ttcaaatgag atggcgaaag tagatgatag tttctttcat cgacttgaag agtctttttt 1801 ggtggaagaa gacaagaagc atgaacgtca tcctattttt ggaaatatag tagatgaagt 1861 tgcttatcat gagaaatatc caactatcta tcatctgcga aaaaaattgg tagattctac 1921 tgataaagcg gatttgcgct taatctattt ggccttagcg catatgatta agtttcgtgg 1981 tcattttttg attgagggag atttaaatcc tgataatagt gatgtggaca aactatttat 2041 ccagttggta caaacctaca atcaattatt tgaagaaaac cctattaacg caagtggagt 2101 agatgctaaa gcgattcttt ctgcacgatt gagtaaatca agacgattag aaaatctcat 2161 tgctcagctc cccggtgaga agaaaaatgg cttatttggg aatctcattg ctttgtcatt 2221 gggtttgacc cctaatttta aatcaaattt tgatttggca gaagatgcta aattacagct 2281 ttcaaaagat acttacgatg atgatttaga taatttattg gcgcaaattg gagatcaata 2341 tgctgatttg tttttggcag ctaagaattt atcagatgct attttacttt cagatatcct 2401 aagagtaaat actgaaataa ctaaggctcc cctatcagct tcaatgatta aacgctacga 2461 tgaacatcat caagacttga ctcttttaaa agctttagtt cgacaacaac ttccagaaaa 2521 gtataaagaa atcttttttg atcaatcaaa aaacggatat gcaggttata ttgatggggg 2581 agctagccaa gaagaatttt ataaatttat caaaccaatt ttagaaaaaa tggatggtac 2641 tgaggaatta ttggtgaaac taaatcgtga agatttgctg cgcaagcaac ggacctttga 2701 caacggctct attccccatc aaattcactt gggtgagctg catgctattt tgagaagaca 2761 agaagacttt tatccatttt taaaagacaa tcgtgagaag attgaaaaaa tcttgacttt 2821 tcgaattcct tattatgttg gtccattggc gcgtggcaat agtcgttttg catggatgac 2881 tcggaagtct gaagaaacaa ttaccccttg gaattttgaa gaagttgtcg ataaaggtgc 2941 ttcagctcaa tcatttattg aacgcatgac aaactttgat aaaaatcttc caaatgaaaa 3001 agtactacca aaacatagtt tgctttatga gtattttacg gtttataacg aattgacaaa 3061 ggtcaaatat gttactgaag gaatgcgaaa accagcattt ctttcaggtg aacagaagaa 3121 agccattgtt gatttactct tcaaaacaaa tcgaaaagta accgttaagc aattaaaaga 3181 agattatttc aaaaaaatag aatgttttga tagtgttgaa atttcaggag ttgaagatag 3241 atttaatgct tcattaggta cctaccatga tttgctaaaa attattaaag ataaagattt 3301 tttggataat gaagaaaatg aagatatctt agaggatatt gttttaacat tgaccttatt 3361 tgaagatagg gagatgattg aggaaagact taaaacatat gctcacctct ttgatgataa 3421 ggtgatgaaa cagcttaaac gtcgccgtta tactggttgg ggacgtttgt ctcgaaaatt 3481 gattaatggt attagggata agcaatctgg caaaacaata ttagattttt tgaaatcaga 3541 tggttttgcc aatcgcaatt ttatgcagct gatccatgat gatagtttga catttaaaga 3601 agacattcaa aaagcacaag tgtctggaca aggcgatagt ttacatgaac atattgcaaa 3661 tttagctggt agccctgcta ttaaaaaagg tattttacag actgtaaaag ttgttgatga 3721 attggtcaaa gtaatggggc ggcataagcc agaaaatatc gttattgaaa tggcacgtga 3781 aaatcagaca actcaaaagg gccagaaaaa ttcgcgagag cgtatgaaac gaatcgaaga 3841 aggtatcaaa gaattaggaa gtcagattct taaagagcat cctgttgaaa atactcaatt 3901 gcaaaatgaa aagctctatc tctattatct ccaaaatgga agagacatgt atgtggacca 3961 agaattagat attaatcgtt taagtgatta tgatgtcgat cacattgttc cacaaagttt 4021 ccttaaagac gattcaatag acaataaggt cttaacgcgt tctgataaaa atcgtggtaa 4081 atcggataac gttccaagtg aagaagtagt caaaaagatg aaaaactatt ggagacaact 4141 tctaaacgcc aagttaatca ctcaacgtaa gtttgataat ttaacgaaag ctgaacgtgg 4201 aggtttgagt gaacttgata aagctggttt tatcaaacgc caattggttg aaactcgcca 4261 aatcactaag catgtggcac aaattttgga tagtcgcatg aatactaaat acgatgaaaa 4321 tgataaactt attcgagagg ttaaagtgat taccttaaaa tctaaattag tttctgactt 4381 ccgaaaagat ttccaattct ataaagtacg tgagattaac aattaccatc atgcccatga 4441 tgcgtatcta aatgccgtcg ttggaactgc tttgattaag aaatatccaa aacttgaatc 4501 ggagtttgtc tatggtgatt ataaagttta tgatgttcgt aaaatgattg ctaagtctga 4561 gcaagaaata ggcaaagcaa ccgcaaaata tttcttttac tctaatatca tgaacttctt 4621 caaaacagaa attacacttg caaatggaga gattcgcaaa cgccctctaa tcgaaactaa 4681 tggggaaact ggagaaattg tctgggataa agggcgagat tttgccacag tgcgcaaagt 4741 attgtccatg ccccaagtca atattgtcaa gaaaacagaa gtacagacag gcggattctc 4801 caaggagtca attttaccaa aaagaaattc ggacaagctt attgctcgta aaaaagactg 4861 ggacccaaaa aaatatggtg gttttgatag tccaacggta gcttattcag tcctagtggt 4921 tgctaaggtg gaaaaaggga aatcgaagaa gttaaaatcc gttaaagagt tactagggat 4981 cacaattatg gaaagaagtt cctttgaaaa aaatccgatt gactttttag aagctaaagg 5041 atataaggaa gttaaaaaag acttaatcat taaactacct aaatatagtc tttttgagtt 5101 agaaaacggt cgtaaacgga tgctggctag tgccggagaa ttacaaaaag gaaatgagct 5161 ggctctgcca agcaaatatg tgaatttttt atatttagct agtcattatg aaaagttgaa 5221 gggtagtcca gaagataacg aacaaaaaca attgtttgtg gagcagcata agcattattt 5281 agatgagatt attgagcaaa tcagtgaatt ttctaagcgt gttattttag cagatgccaa 5341 tttagataaa gttcttagtg catataacaa acatagagac aaaccaatac gtgaacaagc 5401 agaaaatatt attcatttat ttacgttgac gaatcttgga gctcccgctg cttttaaata 5461 ttttgataca acaattgatc gtaaacgata tacgtctaca aaagaagttt tagatgccac 5521 tcttatccat caatccatca ctggtcttta tgaaacacgc attgatttga gtcagctagg 5581 aggtgactca ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg 5641 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc 5701 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa 5761 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 5821 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 5881 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 5941 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 6001 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 6061 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 6121 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 6181 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa 6241 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca 6301 tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca 6361 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg 6421 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca 6481 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca 6541 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt 6601 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc 6661 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa 6721 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct 6781 gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt 6841 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc 6901 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca 6961 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct 7021 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa 7081 tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta 7141 ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt 7201 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 7261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 7321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 7381 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 7441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 7501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 7561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 7621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 7681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 7741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 7801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 7861 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 7921 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 7981 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct 8041 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct 8101 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg 8161 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 8221 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 8281 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt 8341 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg 8401 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg 8461 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga 8521 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta 8581 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc 8641 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga 8701 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga 8761 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc 8821 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta 8881 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga 8941 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga 9001 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa 9061 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga 9121 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc 9181 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct 9241 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 9301 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg 9361 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc 9421 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg 9481 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc 9541 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc 9601 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg 9661 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt 9721 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt 9781 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc 9841 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca 9901 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca 9961 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc 10021 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt 10081 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg 10141 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg 10201 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg 10261 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg 10321 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc 10381 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc 10441 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac 10501 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg 10561 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc 10621 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat 10681 gccggccacg atgcgtccgg cgtagaggat cga //