LOCUS       sequence-401280-        3490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3490)
  AUTHORS   Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
            Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
  TITLE     A scalable framework for high-throughput identification of 
            functional origins of replication in non-model bacteria
  JOURNAL   bioRxiv 2023.05.19.541510
REFERENCE   2  (bases 1 to 3490)
  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: "bioRxiv 
            2023.05.19.541510"
COMMENT     Sequence Label: pGL0_37 [tetA pSC101]
FEATURES             Location/Qualifiers
     source          1..3490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..1272
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(434..453)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(1534..1550)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1534..1550)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1547..1569)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1722..2310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1802..1821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2443..2537)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2558..3217)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3132..3151
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3218..3320)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3407..3429
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3421..3438
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3422..3438
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgaaatct aacaatgcgc tcatcgtcat cctcggcacc
      121 gtcaccctgg atgctgtagg cataggcttg gttatgccgg tactgccggg cctcttgcgg
      181 gatatcgtcc attccgacag catcgccagt cactatggcg tgctgctagc gctatatgcg
      241 ttgatgcaat ttctatgcgc acccgttctc ggagcactgt ccgaccgctt tggccgccgc
      301 ccagtcctgc tcgcttcgct acttggagcc actatcgact acgcgatcat ggcgaccaca
      361 cccgtcctgt ggattctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
      421 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
      481 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
      541 ttgggcgcca tctccctgca cgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
      601 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgccg tccgatgccc
      661 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac cattgtggcc
      721 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
      781 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
      841 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
      901 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
      961 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     1021 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     1081 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcac tggaccgctg
     1141 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
     1201 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     1261 acctcgacct gagctttgag accaccccag gatacataga ttaccacaac tccgagccct
     1321 tccacctagc gtgctaacca cgcacgctat tgttgtatcc catataatgc aatcgtagtc
     1381 tacctgactg tacttagaaa tgtggcttcg cctttgccca cgcacctgat cgctcctcgt
     1441 ttgcttttaa ggaccggacg aaccacagag cattagaagg ctaggtggag gctcagtgat
     1501 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1561 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1621 tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag
     1681 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1741 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1801 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1861 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1921 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1981 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2041 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2101 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2161 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2221 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2281 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2341 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
     2401 aaatcgtccc cccatacgat ataagttgtt actagtgctt ggattctcac caataaaaaa
     2461 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     2521 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     2581 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     2641 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     2701 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     2761 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
     2821 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     2881 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     2941 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     3001 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     3061 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     3121 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     3181 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     3241 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     3301 aacctcttac gtgccgatca acgtctcatt gatacctgaa acaaaaccca tcgtacggcc
     3361 aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt
     3421 tgtaaaacga cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc
     3481 cgcctgacct
//
