LOCUS       sequence-54394-e        3250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3250)
  AUTHORS   Kuhlman TE, Cox EC
  TITLE     Site-specific chromosomal integration of large synthetic constructs.
  JOURNAL   Nucleic Acids Res. 2010 Apr;38(6):e92. Epub 2010 Jan 4.
  PUBMED    20047970
REFERENCE   2  (bases 1 to 3250)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2010-04"; volume: "38(6)"; pages: "e92. Epub 
            2010 Jan 4"
COMMENT     Sequence Label: pTKS/CS
FEATURES             Location/Qualifiers
     source          1..3250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             115..1320
                     /codon_start=1
                     /gene="tetA from transposon Tn10"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNSSTKIALVITLLDAMGIGLIMPVLPTLLREFIASEDIANHFGV
                     LLALYALMQVIFAPWLGKMSDRFGRRPVLLLSLIGASLDYLLLAFSSALWMLYLGRLLS
                     GITGATGAVAASVIADTTSASQRVKWFGWLGASFGLGLIAGPIIGGFAGEISPHSPFFI
                     AALLNIVTFLVVMFWFRETKNTRDNTDTEVGVETQSNSVYITLFKTMPILLIIYFSAQL
                     IGQIPATVWVLFTENRFGWNSMMVGFSLAGLGLLHSVFQAFVAGRIATKWGEKTAVLLG
                     FIADSSAFAFLAFISEGWLVFPVLILLAGGGIALPALQGVMSIQTKSHQQGALQGLLVS
                     LTNATGVIGPLLFAVIYNHSLPIWDGWIWIIGLAFYCIIILLSMTFMLTPQAQGSKQET
                     SA"
     terminator      1369..1455
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1490..1507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1624..2169)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1704..1723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2283..2377)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2398..3057)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2972..2991
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3058..3160)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3226..3244
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tacggcccca aggtccaaac ggtgataggg ataacagggt aatatttacg ttgacaccac
       61 ctttcgcgta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatatgaat
      121 agttcgacaa agatcgcatt ggtaattacg ttactcgatg ccatggggat tggccttatc
      181 atgccagtct tgccaacgtt attacgtgaa tttattgctt cggaagatat cgctaaccac
      241 tttggcgtat tgcttgcact ttatgcgtta atgcaggtta tctttgctcc ttggcttgga
      301 aaaatgtctg accgatttgg tcggcgccca gtgctgttgt tgtcattaat aggcgcatcg
      361 ctggattact tattgctggc tttttcaagt gcgctttgga tgctgtattt aggccgtttg
      421 ctttcaggga tcacaggagc tactggggct gtcgcggcat cggtcattgc cgataccacc
      481 tcagcttctc aacgcgtgaa gtggttcggt tggttagggg caagttttgg gcttggttta
      541 atagcggggc ctattattgg tggttttgca ggagagattt caccgcatag tccctttttt
      601 atcgctgcgt tgctaaatat tgtcactttc cttgtggtta tgttttggtt ccgtgaaacc
      661 aaaaatacac gtgataatac agataccgaa gtaggggttg agacgcaatc aaattcggtg
      721 tacatcactt tatttaaaac gatgcccatt ttgttgatta tttatttttc agcgcaattg
      781 ataggccaaa ttcccgcaac ggtgtgggtg ctatttaccg aaaatcgttt tggatggaat
      841 agcatgatgg ttggcttttc attagcgggt cttggtcttt tacactcagt attccaagcc
      901 tttgtggcag gaagaatagc cactaaatgg ggcgaaaaaa cggcagtact gctcggattt
      961 attgcagata gtagtgcatt tgccttttta gcgtttatat ctgaaggttg gttagttttc
     1021 cctgttttaa ttttattggc tggtggtggg atcgctttac ctgcattaca gggagtgatg
     1081 tctatccaaa caaagagtca tcagcaaggt gctttacagg gattattggt gagccttacc
     1141 aatgcaaccg gtgttattgg cccattactg tttgctgtta tttataatca ttcactacca
     1201 atttgggatg gctggatttg gattattggt ttagcgtttt actgtattat tatcctgcta
     1261 tcaatgacct tcatgttgac ccctcaagct caggggagta aacaggagac aagtgcttag
     1321 tagggataac agggtaatga tggcgcctca tccctgaagc caaggcatca aataaaacga
     1381 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1441 ctgagtagga caaatccgcc gccctagacc tagggatata ttccgcttcc tcgctcactg
     1501 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     1561 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     1621 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     1681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     1741 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     1801 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     1861 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1921 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     1981 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     2041 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     2101 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     2161 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     2221 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt
     2281 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     2341 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
     2401 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2461 gaagccatca cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2521 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2581 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     2641 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2701 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     2761 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     2821 gtctttcatt gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat
     2881 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     2941 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
     3001 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     3061 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     3121 ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccagat
     3181 atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     3241 aggccctttc
//
