LOCUS pDMS197 6199 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDMS197
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6199)
AUTHORS Edwards RA, Keller LH, Schifferli DM
TITLE Improved allelic exchange vectors and their use to analyze 987P
fimbria gene expression.
JOURNAL Gene. 1998 Jan 30;207(2):149-57.
PUBMED 9511756
REFERENCE 2 (bases 1 to 6199)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..6199
/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(3246..4436)
/codon_start=1
/gene="tet"
/product="tetracycline efflux protein"
/label=TcR
/note="confers resistance to tetracycline"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
primer_bind 4065..4084
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter complement(4484..4512)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
oriT 5146..5255
/note="incP origin of transfer"
CDS 5288..5659
/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 ttgggatcca agggttggtt tgcgcattca cagttctccg
3181 caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct
3241 tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca
3301 aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca
3361 taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag agccgcgagc
3421 gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
3481 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
3541 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
3601 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
3661 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
3721 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
3781 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
3841 gggttgaagg ctctcaaggg catcggtcga cgctctccct tatgcgactc ctgcattagg
3901 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
3961 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
4021 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
4081 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
4141 aggatccaca ggacgggtgt ggtcgccatg atcgcgtagt cgatagtggc tccaagtagc
4201 gaagcgagca ggactgggcg gcggccaaag cggtcggaca gtgctccgag aacgggtgcg
4261 catagaaatt gcatcaacgc atatagcgct agcagcacgc catagtgact ggcgatgctg
4321 tcggaatgga cgatatcccg caagaggccc ggcagtaccg gcataaccaa gcctatgcct
4381 acagcatcca gggtgacggt gccgaggatg acgatgagcg cattgttaga tttcatacac
4441 ggtgcctgac tgcgttagca atttaactgt gataaactac cgcattaaag cttatcgatg
4501 ataagctgtc aaacatgaga attgatcctt tttgtccggt gttgggttga aggtgaagcc
4561 ggtcggggcc gcagcggggg ccggcttttc agccttgccc ccctgcttcg gccgccgtgg
4621 ctccggcgtc ttgggtgccg gcgcgggttc cgcagccttg gcctgcggtg cgggcacatc
4681 ggcgggcttg gccttgatgt gccgcctggc gtgcgagcgg aacgtctcgt aggagaactt
4741 gaccttcccc gtttcccgca tgtgctccca aatggtgacg agcgcatagc cggacgctaa
4801 cgccgcctcg acatccgccc tcaccgccag gaacgcaacc gcagcctcat cacgccggcg
4861 cttcttggcc gcgcgggatt caacccactc ggccagctcg tcggtgtagc tctttggcat
4921 cgtctctcgc ctgtcccctc agttcagtaa tttcctgcat ttgcctgttt ccagtcggta
4981 gatattccac aaaacagcag ggaagcagcg cttttccgct gcataaccct gcttcggggt
5041 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
5101 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
5161 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
5221 gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg
5281 caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta
5341 ctgccttcca gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct
5401 gtcggcctac ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga
5461 gcacgtccgc gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa
5521 actctggctc accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct
5581 gctggcgaag atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg
5641 cccgagggca gagccatgac ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg
5701 ccaagcacgt ccccatgcgc tccatcaaga agagcgactt cgcggagctg gtgaagtaca
5761 tcaccgacga gcaaggcaag accgagcgcc tgggtcacgt gcgcgtcacg aactgcgagg
5821 caaacaccct gcccgctgtc atggccgagg tgatggcgac ccagcacggc aacacccgtt
5881 ccgaggccga caagacctat cacctgctgg ttagcttccg cgcgggagag aagcccgacg
5941 cggagacgtt gcgcgcgatt gaggaccgca tctgcgctgg gcttggcttc gccgagcatc
6001 agcgcgtcag tgccgtgcat cacgacaccg acaacctgca catccatatc gccatcaaca
6061 agattcaccc gacccgaaac accatccatg agccgtatcg ggcctaccgc gccctcgctg
6121 acctctgcgc gacgctcgaa cgggactacg ggcttgagcg tgacaatcac gaaacgcggc
6181 agcgcgtttc cgagaaccg
//
