LOCUS Exported 5760 bp ds-DNA circular SYN 21-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5760
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 93..482
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
promoter 510..955
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 956..2377
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
CDS complement(3216..3875)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(3876..3978)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
oriT 4707..4816
/note="incP origin of transfer"
CDS 4849..5220
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
ORIGIN
1 cgcgtttccg agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc
61 ggctggatcg gcacgatgcg tccggcgtag aggatctgaa gatccagcag ttcaacctgt
121 tgatagtacg tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa
181 gctctcatgt ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg
241 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa
301 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa
361 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact
421 gagaagccct tagagcctct caaagcaatt ttcagtgaca caggaacact taacggctga
481 catgggaatt ctgatccttt ttaacccatc acatatacct gccgttcact attatttagt
541 gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc
601 aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta
661 ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca
721 gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt
781 tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca
841 ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag
901 gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa
961 catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg
1021 aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg
1081 catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa
1141 atatcaagtt cctgagttcg attcgtccac aattaaaaat atctcttctg caaaaggcct
1201 ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg
1261 ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta
1321 catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt
1381 ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga
1441 atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt
1501 ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc
1561 agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg
1621 aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa
1681 ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga
1741 agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata
1801 ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa
1861 aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta
1921 cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga
1981 acgcgcgaac gtctttaaaa tgaacggcaa atggtatctg ttcactgact cccgcggatc
2041 aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa
2101 ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct
2161 tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa
2221 caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac
2281 gtttgcgccc agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag
2341 catccttgaa caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgata
2401 tcctattggc attttctttt atttcttatc aacataaagg tgaatcccat atgaactata
2461 taaaagcagg caaatggcta accgtattcc taaccttttg gtaatgactc caacttattg
2521 atagtgtttt atgttcagat aatgcccgat gactttgtca tgcagctcca ccgattttga
2581 gaacgacagc gacttccgtc ccagccgtgc caggtgctgc ctcagattca ggttatgccg
2641 ctcaattcgc tgcgtatatc gcttgctgat tacgtgcagc tttcccttca ggcgggattc
2701 atacagcggc cagccatccg tcatccatat caccacgtca aagggtgaca gcaggctcat
2761 aagacgcccc agcgtcgcca tagtgcgttc accgaatacg tgcgcaacaa ccgtcttccg
2821 gagactgtca tacgcgtaaa acagccagcg ctggcgcgat ttagccccga catagcccca
2881 ctgttcgtcc atttccgcgc agacgatgac gtcactgccc ggctgtatgc gcgaggttac
2941 cgactgcggc ctgagttttt taagtgacgt aaaatcgtgt tgaggccaac gcccataatg
3001 cgggctgttg cccggcatcc aacgccattc atggccatat caatgatttt ctggtgcgta
3061 ccgggttgag aagcggtgta agtgaactgc atgaattccc gggagagctc gatatcgcat
3121 gcggtacctc tagaagaagc ttgggatcgg gccctatcac ttattcaggc gtagcaacca
3181 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
3241 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga
3301 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
3361 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
3421 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
3481 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
3541 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
3601 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
3661 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
3721 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
3781 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
3841 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
3901 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
3961 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
4021 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtattagg
4081 gcccgatcct ttttgtccgg tgttgggttg aaggtgaagc cggtcggggc cgcagcgggg
4141 gccggctttt cagccttgcc cccctgcttc ggccgccgtg gctccggcgt cttgggtgcc
4201 ggcgcgggtt ccgcagcctt ggcctgcggt gcgggcacat cggcgggctt ggccttgatg
4261 tgccgcctgg cgtgcgagcg gaacgtctcg taggagaact tgaccttccc cgtttcccgc
4321 atgtgctccc aaatggtgac gagcgcatag ccggacgcta acgccgcctc gacatccgcc
4381 ctcaccgcca ggaacgcaac cgcagcctca tcacgccggc gcttcttggc cgcgcgggat
4441 tcaacccact cggccagctc gtcggtgtag ctctttggca tcgtctctcg cctgtcccct
4501 cagttcagta atttcctgca tttgcctgtt tccagtcggt agatattcca caaaacagca
4561 gggaagcagc gcttttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
4621 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
4681 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
4741 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
4801 ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat ggctgatgaa
4861 accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc agacgaacga
4921 agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc
4981 gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg cgagctggcc
5041 cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct caccgacgac
5101 ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa gatcgaagag
5161 aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc agagccatga
5221 cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg tccccatgcg
5281 ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg agcaaggcaa
5341 gaccgagcgc ctgggtcacg tgcgcgtcac gaactgcgag gcaaacaccc tgcccgctgt
5401 catggccgag gtgatggcga cccagcacgg caacacccgt tccgaggccg acaagaccta
5461 tcacctgctg gttagcttcc gcgcgggaga gaagcccgac gcggagacgt tgcgcgcgat
5521 tgaggaccgc atctgcgctg ggcttggctt cgccgagcat cagcgcgtca gtgccgtgca
5581 tcacgacacc gacaacctgc acatccatat cgccatcaac aagattcacc cgacccgaaa
5641 caccatccat gagccgtatc gggcctaccg cgccctcgct gacctctgcg cgacgctcga
5701 acgggactac gggcttgagc gtgacaatca cgaaacgcgg cagcgcgttt ccgagaaccg
//