LOCUS       sequence-108473-        4253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derivative of pACYC184 M1uI linkers inserted into EcoRV site 
            inactivating Tc resistance gene. Now Tcs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4253)
  AUTHORS   Donnenberg MS, Calderwood SB, Donohue-Rolfe A, Keusch GT, Kaper JB
  TITLE     Construction and analysis of TnphoA mutants of enteropathogenic 
            Escherichia coli unable to invade HEp-2 cells.
  JOURNAL   Infect Immun. 1990 Jun;58(6):1565-71.
  PUBMED    2160428
REFERENCE   2  (bases 1 to 4253)
  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: "Infect 
            Immun."; date: "1990-06"; volume: "58(6)"; pages: "1565-71"
COMMENT     Sequence Label: pCVD433
FEATURES             Location/Qualifiers
     source          1..4253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /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(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1505..1510,1511..1527,1528..1533)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1941..1960)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(3813..4253,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct
     1561 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg
     1621 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg
     1681 atgacgcgtc atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
     1741 atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
     1801 ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
     1861 gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     1921 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     1981 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     2041 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     2101 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
     2161 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2221 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2281 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     2341 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     2401 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     2461 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     2521 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     2581 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcactgg
     2641 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     2701 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     2761 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     2821 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     2881 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     2941 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     3001 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     3061 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     3121 tcgtaaagtc tggaaacgcg gaagtcccct acgtgctgct gaagttgccc gcaacagaga
     3181 gtggaaccaa ccggtgatac cacgatacta tgactgagag tcaacgccat gagcggcctc
     3241 atttcttatt ctgagttaca acagtccgca ccgctgtccg gtagctcctt ccggtgggcg
     3301 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga
     3361 caggtgccgg cagcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     3421 aaacaagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
     3481 tgtggaacac ctacatctgt attaacgaag cgctaaccgt ttttatcagg ctctgggagg
     3541 cagaataaat gatcatatcg tcaattatta cctccacggg gagagcctga gcaaactggc
     3601 ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc
     3661 agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt
     3721 tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt agcaccaggc
     3781 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     3841 tactgttgta attcattaag cattctgccg acatggaagc catcacagac ggcatgatga
     3901 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     3961 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     4021 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     4081 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     4141 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     4201 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acg
//
