LOCUS       sequence-385365-        3909 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 3909)
  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 3909)
  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_7 [pSa ORI]
FEATURES             Location/Qualifiers
     source          1..3909
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(386..480)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(501..1160)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1075..1094
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1161..1263)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1350..1372
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1364..1381
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1365..1381
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1799..2770
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein of plasmid pSa"
                     /label=RepA
                     /translation="MPKNNKAPGHRINEIIKTSLALEMEDAREAGLVGYMARCLVQATM
                     PHTDPKTSYFERTNGIVTLSIMGKPSIGLPYGSMPRTLLAWICTEAVRTKDPVLNLGRS
                     QSEFLQRLGMHTDGRYTATLRNQAQRLFSSMISLAGEQGNDFGIENVVIAKRAFLFWNP
                     KRPEDRALWDSTLTLTGDFFEEVTRSPVPIRIDYLHALRQSPLAMDIYTWLTYRVFLLR
                     AKGRPFVQIPWVALQAQFGSSYGSRARNSPELDDKARERAERAALASFKYNFKKRLREV
                     LIVYPEASDCIEDDGECLRIKSTRLHVTRAPGKGARIGPPPT"
     rep_origin      2807..3242
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(3386..3402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3386..3402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3399..3421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(3574..3909,1..253))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3654..3673)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
       61 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      121 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      181 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      241 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc tattcaacaa
      301 agccgccgtc ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac
      361 gatataagtt gttactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
      421 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
      481 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
      541 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
      601 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
      661 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
      721 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
      781 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
      841 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
      901 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
      961 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     1021 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     1081 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     1141 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     1201 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     1261 tcaacgtctc attgatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
     1321 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1381 tcatgcataa tccgcacgca tctggaataa ggaagtgcca ttccgcctga ccttagcgtg
     1441 cgaacagcac gctattgttg tatcattact cgcatccatt ctcaggctgt ctcgtctcgt
     1501 ctccgaggtc tccggaggat cgagcggcat ttcattcagg cacgtaccac cgaggcgcta
     1561 caagtcgcca gagagcgcgg caagacgctc gggcgaccca agggcagcaa atcgagcgcc
     1621 ttgaagctgg acagccgtat tgatgaagta caggcatacg tgaaccttgg cttgccgcaa
     1681 agtcgcgcag ccgagttgtt aggcgtcagc cctcacacct tgcgcctgtt catcaaacgc
     1741 cggaacatca aacccacaaa cactagacca accatcacca tgccggggag ggaacaacat
     1801 gcctaagaac aacaaagccc ccggccatcg tatcaacgag atcatcaaga cgagcctcgc
     1861 gctcgaaatg gaggatgccc gcgaagctgg cttagtcggc tacatggccc gttgccttgt
     1921 gcaagcgacc atgccccaca ccgaccccaa gaccagctac tttgagcgca ccaatggcat
     1981 cgtcaccttg tcgatcatgg gcaagccgag catcggcctg ccctacggtt ctatgccgcg
     2041 caccttgctt gcttggatat gcaccgaggc cgtgcgaacg aaagaccccg tgttgaacct
     2101 tggccggtcg caatcggaat ttctacaaag gctcggaatg cacaccgatg gccgttacac
     2161 ggccaccctt cgcaatcagg cgcaacgcct gttttcatcc atgatttcgc ttgccggcga
     2221 gcaaggcaat gacttcggca ttgagaacgt cgtcattgcc aagcgcgctt ttctattctg
     2281 gaatcccaag cggccagaag atcgggcgct atgggatagc accctcaccc tcacaggcga
     2341 tttcttcgag gaagtcaccc gctcaccggt tcctatccga atcgactacc tgcatgcctt
     2401 gcggcagtct ccgcttgcga tggacattta cacgtggctg acctatcgcg tgttcctgtt
     2461 gcgggccaag ggccgcccct tcgtgcaaat cccttgggtc gccctgcaag cgcaattcgg
     2521 ctcatcctat ggcagccgcg cacgcaactc gcccgaactg gacgataagg cccgagagcg
     2581 ggcagagcgg gcagcactcg ccagcttcaa atacaacttc aaaaagcgcc tacgcgaagt
     2641 gttgattgtc tatcccgagg caagcgactg catcgaagat gacggcgaat gcctgcgcat
     2701 caaatccaca cgcctgcatg tcacccgcgc acccggcaag ggcgctcgca tcggcccccc
     2761 tccgacttga ccaggccaac gctacgcttg gcttggtcaa gccttcccat ccaacagccc
     2821 gccgtcgagc gggctttttt atccccggaa gcctgtggat agagggtagt tatccacgtg
     2881 aaaccgctaa tgccccgcaa agccttgatt cacggggctt tccggcccgc tccaaaaact
     2941 atccacgtga aatcgctaat cagggtacgt gaaatcgcta atcggagtac gtgaaatcgc
     3001 taataaggtc acgtgaaatc gctaatcaaa aaggcacgtg agaacgctaa tagccctttc
     3061 agatcaacag cttgcaaaca cccctcgctc cggcaagtag ttacagcaag tagtatgttc
     3121 aattagcttt tcaattatga atatatatat caattattgg tcgcccttgg cttgtggaca
     3181 atgcgctacg cgcaccggct ccgcccgtgg acaaccgcaa gcggttgccc accgtcgagc
     3241 gcccgctcga gaccacccca ggatacatag attaccacaa ctccgagccc ttccacctag
     3301 cgtgctaacc acgcacgcta ttgttgtata ggctaggtgg aggctcagtg atgataagtc
     3361 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3421 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
     3481 agttcggcgt atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3541 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3601 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3661 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3721 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3781 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3841 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3901 acacgactt
//
