LOCUS       sequence-427292-        3452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium copy thyA-infA plasmid containing spectinomycin as antibiotic
            resistance gene (ARG) for validation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3452)
  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 3452)
  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: pSV013
FEATURES             Location/Qualifiers
     source          1..3452
                     /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"
     terminator      complement(1229..1323)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1703..2494
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPVQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      2526..2597
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2613..2640
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2668..2685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2802..3347)
                     /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(2882..2901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgacagcta gctcagtcct aggtataatg ctagctcgac cgctatttta gcccatatca
       61 gaggtaacac ctaatgaaac agtatttaga actgatgcaa aaagtgctcg acgaaggcac
      121 acagaaaaac gaccgtaccg gaaccggaac gctttccatt tttggtcatc agatgcgttt
      181 taacctgcaa gatggattcc cgctggtgac aactaaacgt tgccacctgc gttccatcat
      241 ccatgaactg ctgtggtttc tgcagggcga cactaacatt gcttatctac acgaaaacaa
      301 tgtcaccatc tgggacgaat gggccgatga aaacggcgac ctcgggccag tgtatggtaa
      361 acagtggcgc gcctggccaa cgccagatgg tcgtcatatt gaccagatca ctacggtact
      421 gaaccagctg aaaaacgacc cggattcgcg ccgcattatt gtttcagcgt ggaacgtagg
      481 cgaactggat aaaatggcgc tggcaccgtg ccatgcattc ttccagttct atgtggcaga
      541 cggcaaactc tcttgccagc tttatcagcg ctcctgtgac gtcttcctcg gcctgccgtt
      601 caacattgcc agctacgcgt tattggtgca tatgatggcg cagcagtgcg atctggaagt
      661 gggtgatttt gtctggaccg gtggcgacac gcatctgtac agcaaccata tggatcaaac
      721 tcatctgcaa ttaagccgcg aaccgcgtcc gctgccgaag ttgattatca aacgtaaacc
      781 cgaatccatc ttcgactacc gtttcgaaga ctttgagatt gaaggctacg atccgcatcc
      841 gggcattaaa gcgccggtgg ctatctaagg ttcaaattac ggtagtgata ccccagagga
      901 ttagatggcc aaagaagaca atattgaaat gcaaggtacc gttcttgaaa cgttgcctaa
      961 taccatgttc cgcgtagagt tagaaaacgg tcacgtggtt actgcacaca tctccggtaa
     1021 aatgcgcaaa aactacatcc gcatcctgac gggcgacaaa gtgactgttg aactgacccc
     1081 gtacgacctg agcaaaggcc gcattgtctt ccgtagtcgc tgaagcctca aagtaatcaa
     1141 gccctggtaa ctgccagggc ttttttattt catctttgaa tcaacatatt aaaattcatc
     1201 gaatggcatc cttgctaaca gtgcttgtat tctcaccaat aaaaaacgcc cggcggcaac
     1261 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     1321 gtccaagcga gctctcgaac cccagagtcc cagccaggac agaaatgcct cgacttcgct
     1381 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     1441 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     1501 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     1561 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     1621 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     1681 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     1741 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     1801 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     1861 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     1921 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     1981 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     2041 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     2101 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     2161 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     2221 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     2281 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     2341 ggagcgcctg ccggtccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     2401 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     2461 gaaaggcgag atcaccaagg tagtcggcaa ataagatcca aactcgagta aggatctcca
     2521 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2581 tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg
     2641 tttataccta gggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     2701 actgcggcga gcgaaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     2761 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     2821 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     2881 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     2941 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     3001 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     3061 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     3121 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     3181 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     3241 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     3301 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     3361 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcagagg
     3421 ctagtgtaga tcgctactag agcgcactga cc
//
