LOCUS       sequence-427293-        3639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy thyA-infA plasmid containing chloramphenicol as antibiotic
            resistance gene (ARG) for validation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3639)
  AUTHORS   Amrofell MB, Rengarajan S, Vo ST, Ramirez Tovar ES, LoBello L, 
            Dantas G, Moon TS
  TITLE     Engineering E. coli strains using antibiotic-resistance-gene-free 
            plasmids.
  JOURNAL   Cell Rep Methods. 2023 Dec 18;3(12):100669. doi: 
            10.1016/j.crmeth.2023.100669. Epub 2023 Dec 11.
  PUBMED    38086386
REFERENCE   2  (bases 1 to 3639)
  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: "Cell Rep 
            Methods."; date: "2023-12-18"; volume: "3(12):100669. doi"; pages: "
            10.1016/j.crmeth.2023.100669. Epub 2023 Dec 11"
COMMENT     Sequence Label: pSV014
FEATURES             Location/Qualifiers
     source          1..3639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     primer_bind     complement(1235..1252)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1406..1994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1486..1505)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2165..2824)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTIGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2739..2758
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2825..2927)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3028..3046
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctgacagcta gctcagtcct aggtataatg ctagctcgac cgctatttta gcatcagagg
       61 taacacctaa tgaaacagta tttagaactg atgcaaaaag tgctcgacga aggcacacag
      121 aaaaacgacc gtaccggaac cggaacgctt tccatttttg gtcatcagat gcgttttaac
      181 ctgcaagatg gattcccgct ggtgacaact aaacgttgcc acctgcgttc catcatccat
      241 gaactgctgt ggtttctgca gggcgacact aacattgctt atctacacga aaacaatgtc
      301 accatctggg acgaatgggc cgatgaaaac ggcgacctcg ggccagtgta tggtaaacag
      361 tggcgcgcct ggccaacgcc agatggtcgt catattgacc agatcactac ggtactgaac
      421 cagctgaaaa acgacccgga ttcgcgccgc attattgttt cagcgtggaa cgtaggcgaa
      481 ctggataaaa tggcgctggc accgtgccat gcattcttcc agttctatgt ggcagacggc
      541 aaactctctt gccagcttta tcagcgctcc tgtgacgtct tcctcggcct gccgttcaac
      601 attgccagct acgcgttatt ggtgcatatg atggcgcagc agtgcgatct ggaagtgggt
      661 gattttgtct ggaccggtgg cgacacgcat ctgtacagca accatatgga tcaaactcat
      721 ctgcaattaa gccgcgaacc gcgtccgctg ccgaagttga ttatcaaacg taaacccgaa
      781 tccatcttcg actaccgttt cgaagacttt gagattgaag gctacgatcc gcatccgggc
      841 attaaagcgc cggtggctat ctaaggttca aattacggta gtgatacccc agaggattag
      901 atggccaaag aagacaatat tgaaatgcaa ggtaccgttc ttgaaacgtt gcctaatacc
      961 atgttccgcg tagagttaga aaacggtcac gtggttactg cacacatctc cggtaaaatg
     1021 cgcaaaaact acatccgcat cctgacgggc gacaaagtga ctgttgaact gaccccgtac
     1081 gacctgagca aaggccgcat tgtcttccgt agtcgctgaa gccgagcaag acaatggtaa
     1141 gtagtaatag ctcctgcccc cggaagatca ttccgggggc ttttttattc ccactgtgaa
     1201 ctggatcctt actcgagtct agactgcagg cttcctcgct cactgactcg ctgcgctcgg
     1261 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1321 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1381 gtaaaaaggc cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca
     1441 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1501 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1561 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1621 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1681 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1741 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1801 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1861 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccgcca
     1921 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1981 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2041 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2101 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2161 acagttacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     2221 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     2281 tgtcgccttg cgtataatat ttgccaatgg tgaaaacggg ggcgaagaag ttgtccatat
     2341 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     2401 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     2461 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2521 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2581 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     2641 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     2701 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2761 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2821 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     2881 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     2941 gccagatatc gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3001 attattatca tgacattaac ctataaaaat aggcgtatca cgaggcatca ccctaggtgt
     3061 aaaacgacgg ccagactcgg taccaaattc cagaaaagag acgctgaaaa gcgtcttttt
     3121 tcgttttggt ccttattaat ttttaaagta tgggcttaac cagattagat cccgtagatg
     3181 tccatccccg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     3241 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     3301 ctctccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     3361 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     3421 gttgaaggct ctcaagggca tcggtcgaga tccgggctta tcgactgcac ggtgcaccaa
     3481 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
     3541 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
     3601 tcataacggt tctggcaaat attctgaaat gagctgacc
//
