LOCUS Exported 9275 bp ds-DNA circular SYN 26-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..9275
/organism="synthetic DNA construct"
/mol_type="other DNA"
oriT 618..727
/note="incP origin of transfer"
CDS 760..1131
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
rep_origin 1258..1969
/label=oriV
/note="incP origin of replication"
promoter complement(2505..2523)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2541..2557)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2565..2581
/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)."
promoter complement(2589..2619)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2634..2655
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2902..2930
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 2978..4168
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
CDS 4667..5815
/codon_start=1
/product="trans-acting replication protein that binds to
and activates oriV"
/label=trfA
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
CDS 7196..8167
/codon_start=1
/gene="repA"
/product="replication protein of plasmid pSa"
/label=RepA
/translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
CDS 8562..9212
/codon_start=1
/gene="tetR"
/product="tetracycline resistance regulatory protein"
/label=TetR
/translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
ORIGIN
1 cccctttcca ccgcgtcaga cgcccgtagc agcccgctac gggctttttc atgccctgcc
61 ctagcgtcca agcctcacgg ccgcgctcgg cctctctggc ggccttctgg cgctcttccg
121 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
181 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
241 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
301 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
361 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
421 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
481 cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
541 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
601 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
661 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
721 tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca
781 accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg aagagcgatt
841 gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc cgtcggccag
901 ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc ccgcatcaat
961 ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga cccgcgcacg
1021 gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga gaagcaggac
1081 gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg acttttttag
1141 ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc gctccatcaa
1201 gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca agaccgagcg
1261 cctttgcgac gctcaccggg ctggttgccc tcgccgctgg gctggcggcc gtctatggcc
1321 ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccgcggc cgccggcgtt
1381 gtggatacct cgcggaaaac ttggccctca ctgacagatg aggggcggac gttgacactt
1441 gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga tttcggccgg
1501 cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc ctgattttac gcgagtttcc
1561 cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac ttgaggggcg
1621 cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt gctgacagat
1681 gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc agcatttgca
1741 agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct tttaaaccaa
1801 tatttataaa ccttgttttt aaccagggct gcgccctgtg cgcgtgaccg cgcacgccga
1861 aggggggtgc ccccccttct cgaaccctcc cggcccgcta acgcgggcct cccatccccc
1921 caggggctgc gcccctcggc cgcgaacggc ctcaccccaa aaatggcagc gctggcagtc
1981 cttgccattg ccgggatcgg ggcagtaacg ggatgggcga tcagcccgag cgcgacgccc
2041 ggaagcattg acgtgccgca ggtgctggca tcgacattca gcgaccaggt gccgggcagt
2101 gagggcggcg gcctgggtgg cggcctgccc ttcacttcgg ccgtcggggc attcacggac
2161 ttcatggcgg ggccggcaat ttttaccttg ggcattcttg gcatagtggt cgcgggtgcc
2221 gtgctcgtgt tcgggggtgc gataaaccca gcgaaccatt tgaggtgata ggtaagatta
2281 taccgaggta tgaaaacgag aattggacct ttacagaatt actctatgaa gcgccatatt
2341 taaaaagcta ccaagacgaa gaggatgaag aggatgagga ggcagattgc cttgaatata
2401 ttgacaatac tgataagata atatatcttt tatatagaag ataattcgag ctcggtaccc
2461 ggggatcctc tagagtcgac ctgcaggcat gcaagcttga gtattctata gtgtcaccta
2521 aatagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
2581 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
2641 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
2701 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
2761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
2821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
2881 aacatgaatt aattctcatg tttgacagct tatcatcgat tagctttaat gcggtagttt
2941 atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat
3001 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact
3061 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct
3121 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga
3181 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc
3241 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat
3301 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
3361 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
3421 cgtggccggg ggactgttgg gcgccatctc cttacatgca ccattccttg cggcggcggt
3481 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
3541 gcgccgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
3601 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
3661 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
3721 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
3781 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
3841 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
3901 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
3961 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
4021 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
4081 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
4141 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
4201 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
4261 ccttgtatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
4321 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta cctagatgtg
4381 gcgcaacgat gccggcgaca agcaggagcg caccgacttc ttccgcatca agtgttttgg
4441 ctctcaggcc gaggcccacg gcaagtattt gggcaagggg tcgctggtat tcgtgcaggg
4501 caagattcgg aataccaagt acgagaagga cggccagacg gtctacggga ccgacttcat
4561 tgccgataag gtggattatc tggacaccaa ggcaccaggc gggtcaaatc aggaataagg
4621 gcacattgcc ccggcgtgag tcggggcaat cccgcaagga gggtgaatga atcggacgtt
4681 tgaccggaag gcatacaggc aagaactgat cgacgcgggg ttttccgccg aggatgccga
4741 aaccatcgca agccgcaccg tcatgcgtgc gccccgcgaa accttccagt ccgtcggctc
4801 gatggtccag caagctacgg ccaagatcga gcgcgacagc gtgcaactgg ctccccctgc
4861 cctgcccgcg ccatcggccg ccgtggagcg ttcgcgtcgt ctcgaacagg aggcggcagg
4921 tttggcgaag tcgatgacca tcgacacgcg aggaactatg acgaccaaga agcgaaaaac
4981 cgccggcgag gacctggcaa aacaggtcag cgaggccaag caggccgcgt tgctgaaaca
5041 cacgaagcag cagatcaagg aaatgcagct ttccttgttc gatattgcgc cgtggccgga
5101 cacgatgcga gcgatgccaa acgacacggc ccgctctgcc ctgttcacca cgcgcaacaa
5161 gaaaatcccg cgcgaggcgc tgcaaaacaa ggtcattttc cacgtcaaca aggacgtgaa
5221 gatcacctac accggcgtcg agctgcgggc cgacgatgac gaactggtgt ggcagcaggt
5281 gttggagtac gcgaagcgca cccctatcgg cgagccgatc accttcacgt tctacgagct
5341 ttgccaggac ctgggctggt cgatcaatgg ccggtattac acgaaggccg aggaatgcct
5401 gtcgcgccta caggcgacgg cgatgggctt cacgtccgac cgcgttgggc acctggaatc
5461 ggtgtcgctg ctgcaccgct tccgcgtcct ggaccgtggc aagaaaacgt cccgttgcca
5521 ggtcctgatc gacgaggaaa tcgtcgtgct gtttgctggc gaccactaca cgaaattcat
5581 atgggagaag taccgcaagc tgtcgccgac ggcccgacgg atgttcgact atttcagctc
5641 gcaccgggag ccgtacccgc tcaagctgga aaccttccgc ctcatgtgcg gatcggattc
5701 cacccgcgtg aagaagtggc gcgagcaggt cggcgaagcc tgcgaagagt tgcgaggcag
5761 cggcctggtg gaacacgcct gggtcaatga tgacctggtg cattgcaaac gctagggcct
5821 tgtggggtca gttccggctg ggggttcagc agccagcgct ttactggcat ttcaggaaca
5881 agcgggcact gctcgacgca cttgcttcgc tcagtatcgc tcgggacgca cggcgcgctc
5941 tacgaactgc cgataaacag aggattaaaa ttgacaattg tgattaaggc tcagattcga
6001 cggcttggag cggccgacgt gcaggatttc cgcgagatcc gattgtcggc cctgaagaaa
6061 gctccagaga tgttcgggtc cgtttacgag cacgaggaga aaaagcccat ggaggcgttc
6121 gctgaacggt tgcgagatgc cgtggcattc ggcgcctaca tcgacggcga gatcattggg
6181 ctgtcggtct tcaaacagga ggacggcccc aaggacgctc acaaggcgca tctgtccggc
6241 gttttcgtgg agcccgaaca gcgaggccga ggggtcgccg gtatgctgct gcgggcgttg
6301 ccggcgggtt tattgctcgt gatgatcgtc cgacagattc caacgggaat ctggtggatg
6361 cgcatcttca tcctcggcgc acttaatatt tcgctattct ggagcttgtt gtttatttcg
6421 gtctaccgcc tgccgggcgg ggtcgcggcg acggtaggcg ctgtgcagcc gctgatggtc
6481 gtgttcatct ctgccgctct gctaggtagc ccgatacgat tgatggcggt cctgggggct
6541 atttgcggaa ctgcgggcgt ggcgctgttg gtgttgacac caaacgcagc gctagatcct
6601 gtcggcgtcg cagcgggcct ggcgggggcg gtttccatgg cgttcggaac cgtgctgacc
6661 cgcaagtggc aacctcccgt gcctctgctc acctttaccg cctggcaact ggcggccgga
6721 ggacttctgc tcgttccagt agctttagtg tttgatccgc caatcccgat gcctacagga
6781 accaatgttc tcggcctggc gtggctcggc ctgatcggag cgggtttaac ctacttcctt
6841 tggttccggg ggatctcgcg actcgaacct acagttgttt ccttactggg ctttctcagc
6901 cccagatcct agaggatcga gcggcatttc attcaggcac gtaccaccga ggcgctacaa
6961 gtcgccagag agcgcggcaa gacgctcggg cgacccaagg gcagcaaatc gagcgccttg
7021 aagctggaca gccgtattga tgaagtacag gcatacgtga accttggctt gccgcaaagt
7081 cgcgcagccg agttgttagg cgtcagccct cacaccttgc gcctgttcat caaacgccgg
7141 aacatcaaac ccacaaacac tagaccaacc atcaccatgc cggggaggga acaacatgcc
7201 taagaacaac aaagcccccg gccatcgtat caacgagatc atcaagacga gcctcgcgct
7261 cgaaatggag gatgcccgcg aagctggctt agtcggctac atggcccgtt gccttgtgca
7321 agcgaccatg ccccacaccg accccaagac cagctacttt gagcgcacca atggcatcgt
7381 caccttgtcg atcatgggca agccgagcat cggcctgccc tacggttcta tgccgcgcac
7441 cttgcttgct tggatatgca ccgaggccgt gcgaacgaaa gaccccgtgt tgaaccttgg
7501 ccggtcgcaa tcggaatttc tacaaaggct cggaatgcac accgatggcc gttacacggc
7561 cacccttcgc aatcaggcgc aacgcctgtt ttcatccatg atttcgcttg ccggcgagca
7621 aggcaatgac ttcggcattg agaacgtcgt cattgccaag cgcgcttttc tattctggaa
7681 tcccaagcgg ccagaagatc gggcgctatg ggatagcacc ctcaccctca caggcgattt
7741 cttcgaggaa gtcacccgct caccggttcc tatccgaatc gactacctgc acgccttgcg
7801 gcagtctccg cttgcgatgg acatttacac gtggctgacc tatcgcgtgt tcctgttgcg
7861 ggccaagggc cgccccttcg tgcaaatccc ttgggtcgcc ctgcaagcgc aattcggctc
7921 atcctatggc agccgcgcac gcaactcgcc cgaactggac gataaggccc gagagcgggc
7981 agagcgggca gcactcgcca gcttcaaata caacttcaaa aagcgcctac gcgaagtgtt
8041 gattgtctat cccgaggcaa gcgactgcat cgaagatgac ggcgaatgcc tgcgcatcaa
8101 atccacacgc ctgcatgtca cccgcgcacc cggcaagggc gctcgcatcg gcccccctcc
8161 gacttgacca ggccaacgct acgcttggct tggtcaagga tcgatctggc gccggccagc
8221 gagacgagca agattggccg ccgcccgaaa cgatccgaca gcgcgcccag cacaggtgcg
8281 caggcaaatt gcaccaacgc atacagcgcc agcagaatgc catagtgggc ggtgacgtcg
8341 ttcgagtgaa ccagatcgcg caggaggccc ggcagcaccg gcataatcag gccgatgccg
8401 acagcgtcga gcgcgacagt gctcagaatt acgatcaggg gtatgttggg tttcacgtct
8461 ggcctccgga ccagcctccg ctggtccgat tgaacgcgcg gattctttat cactgataag
8521 ttggtggaca tattatgttt atcagtgata aagtgtcaag catgacaaag ttgcagccga
8581 atacagtgat ccgtgccgcc ctggacctgt tgaacgaggt cggcgtagac ggtctgacga
8641 cacgcaaact ggcggaacgg ttgggggttc agcagccggc gctttactgg cacttcagga
8701 acaagcgggc gctgctcgac gcactggccg aagccatgct ggcggagaat catacgcatt
8761 cggtgccgag agccgacgac gactggcgct catttctgat cgggaatgcc cgcagcttca
8821 ggcaggcgct gctcgcctac cgcgatggcg cgcgcatcca tgccggcacg cgaccgggcg
8881 caccgcagat ggaaacggcc gacgcgcagc ttcgcttcct ctgcgaggcg ggtttttcgg
8941 ccggggacgc cgtcaatgcg ctgatgacaa tcagctactt cactgttggg gccgtgcttg
9001 aggagcaggc cggcgacagc gatgccggcg agcgcggcgg caccgttgaa caggctccgc
9061 tctcgccgct gttgcgggcc gcgatagacg ccttcgacga agccggtccg gacgcagcgt
9121 tcgagcaggg actcgcggtg attgtcgatg gattggcgaa aaggaggctc gttgtcagga
9181 acgttgaagg accgagaaag ggtgacgatt gatcaggacc gctgccggag cgcaacccac
9241 tcactacagc agagccatgt agacaacatc ccctc
//