LOCUS Exported 5286 bp ds-DNA circular SYN 08-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5286
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 4..392
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(1010..1381)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 1414..1523
/note="incP origin of transfer"
CDS complement(2489..3148)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS complement(3246..4667)
/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"
promoter complement(4668..5113)
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
protein_bind 5239..5255
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
1 ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc
61 ttctcagtgc gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa
121 aagccttata tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg
181 ctgatttata ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct
241 tagtacgtta gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg
301 agagcttagt acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac
361 tatcaacagg ttgaactgct gatcttcaga tcctctacgc cggacgcatc gtggccggat
421 ccagccgacc aggctttcca cgcccgcgtg ccgctccatg tcgttcgcgc ggttctcgga
481 aacgcgctgc cgcgtttcgt gattgtcacg ctcaagcccg tagtcccgtt cgagcgtcgc
541 gcagaggtca gcgagggcgc ggtaggcccg atacggctca tggatggtgt ttcgggtcgg
601 gtgaatcttg ttgatggcga tatggatgtg caggttgtcg gtgtcgtgat gcacggcact
661 gacgcgctga tgctcggcga agccaagccc agcgcagatg cggtcctcaa tcgcgcgcaa
721 cgtctccgcg tcgggcttct ctcccgcgcg gaagctaacc agcaggtgat aggtcttgtc
781 ggcctcggaa cgggtgttgc cgtgctgggt cgccatcacc tcggccatga cagcgggcag
841 ggtgtttgcc tcgcagttcg tgacgcgcac gtgacccagg cgctcggtct tgccttgctc
901 gtcggtgatg tacttcacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
961 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
1021 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
1081 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
1141 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
1201 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
1261 aggccgacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
1321 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
1381 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
1441 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
1501 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
1561 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
1621 ctataatgac cccgaagcag ggttatgcag cggaaaagcg ctgcttccct gctgttttgt
1681 ggaatatcta ccgactggaa acaggcaaat gcaggaaatt actgaactga ggggacaggc
1741 gagagacgat gccaaagagc tacaccgacg agctggccga gtgggttgaa tcccgcgcgg
1801 ccaagaagcg ccggcgtgat gaggctgcgg ttgcgttcct ggcggtgagg gcggatgtcg
1861 aggcggcgtt agcgtccggc tatgcgctcg tcaccatttg ggagcacatg cgggaaacgg
1921 ggaaggtcaa gttctcctac gagacgttcc gctcgcacgc caggcggcac atcaaggcca
1981 agcccgccga tgtgcccgca ccgcaggcca aggctgcgga acccgcgccg gcacccaaga
2041 cgccggagcc acggcggccg aagcaggggg gcaaggctga aaagccggcc cccgctgcgg
2101 ccccgaccgg cttcaccttc aacccaacac cggacaaaaa ggatcctcta cgccggacgc
2161 atcgtggccg gcatcaccgg cgccacaagt gcggttgctg gcgcctatat cgccgacatc
2221 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
2281 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgct gcctcgcgcg
2341 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
2401 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
2461 gtgtcggggc gcagccatga cccgggaatt acgccccgcc ctgccactca tcgcagtact
2521 gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct
2581 gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa
2641 cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc
2701 agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt
2761 tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt
2821 ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag
2881 ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat
2941 gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt
3001 tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
3061 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
3121 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgcc ctatgggatt
3181 cacctttatg ttgataagaa ataaaagaaa atgccaatag gatatcggca ttttcttttg
3241 cgtttttatt tgttaactgt taattgtcct tgttcaagga tgctgtcttt gacaacagat
3301 gttttcttgc ctttgatgtt cagcaggaag cttggcgcaa acgttgattg tttgtctgcg
3361 tagaatcctc tgtttgtcat atagcttgta atcacgacat tgtttccttt cgcttgaggt
3421 acagcgaagt gtgagtaagt aaaggttaca tcgttaggat caagatccat ttttaacaca
3481 aggccagttt tgttcagcgg cttgtatggg ccagttaaag aattagaaac ataaccaagc
3541 atgtaaatat cgttagacgt aatgccgtca atcgtcattt ttgatccgcg ggagtcagtg
3601 aacaggtacc atttgccgtt cattttaaag acgttcgcgc gttcaatttc atctgttact
3661 gtgttagatg caatcagcgg tttcatcact tttttcagtg tgtaatcatc gtttagctca
3721 atcataccga gagcgccgtt tgctaactca gccgtgcgtt ttttatcgct ttgcagaagt
3781 ttttgacttt cttgacggaa gaatgatgtg cttttgccat agtatgcttt gttaaataaa
3841 gattcttcgc cttggtagcc atcttcagtt ccagtgtttg cttcaaatac taagtatttg
3901 tggcctttat cttctacgta gtgaggatct ctcagcgtat ggttgtcgcc tgagctgtag
3961 ttgccttcat cgatgaactg ctgtacattt tgatacgttt ttccgtcacc gtcaaagatt
4021 gatttataat cctctacacc gttgatgttc aaagagctgt ctgatgctga tacgttaact
4081 tgtgcagttg tcagtgtttg tttgccgtaa tgtttaccgg agaaatcagt gtagaataaa
4141 cggatttttc cgtcagatgt aaatgtggct gaacctgacc attcttgtgt ttggtctttt
4201 aggatagaat catttgcatc gaatttgtcg ctgtctttaa agacgcggcc agcgtttttc
4261 cagctgtcaa tagaagtttc gccgactttt tgatagaaca tgtaaatcga tgtgtcatcc
4321 gcatttttag gatctccggc taatgcaaag acgatgtggt agccgtgata gtttgcgaca
4381 gtgccgtcag cgttttgtaa tggccagctg tcccaaacgt ccaggccttt tgcagaagag
4441 atatttttaa ttgtggacga atcgaattca ggaacttgat atttttcatt tttttgctgt
4501 tcagggattt gcagcatatc atggcgtgta atatgggaaa tgccgtatgt ttccttatat
4561 ggcttttggt tcgtttcttt cgcaaacgct tgagttgcgc ctcctgccag cagtgcggta
4621 gtaaaggtta atactgttgc ttgttttgca aactttttga tgttcatcgt tcatgtctcc
4681 ttttttatgt actgtgttag cggtctgctt cttccagccc tcctgtttga agatggcaag
4741 ttagttacgc acaataaaaa aagacctaaa atatgtaagg ggtgacgcca aagtatacac
4801 tttgcccttt acacatttta ggtcttgcct gctttatcag taacaaaccc gcgcgattta
4861 cttttcgacc tcattctatt agactctcgt ttggattgca actggtctat tttcctcttt
4921 tgtttgatag aaaatcataa aaggatttgc agactacggg cctaaagaac taaaaaatct
4981 atctgtttct tttcattctc tgtatttttt atagtttctg ttgcatgggc ataaagttgc
5041 ctttttaatc acaattcaga aaatatcata atatctcatt tcactaaata atagtgaacg
5101 gcaggtatat gtgatgggtt aaaaaggatc gatcctctag agtcgacctg caggtcgacg
5161 gatctgcagg tcgacggatc ccaagcttct tctagaggta ccgcatgcga tatcgagctc
5221 tcccgggaat tccacaaatt gttatccgct cacaattcca catgtggaat tccacatgtg
5281 gaattc
//