LOCUS       sequence-393042-        4052 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 4052)
  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 4052)
  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_5 [pAL5000 ORI]
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2096..2112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2096..2112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2109..2131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2284..2872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2364..2383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3005..3099)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3120..3779)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3694..3713
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3780..3882)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3969..3991
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3983..4000
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3984..4000
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagagcgtc taccaggact tttacctgtc cgacccgttg
      121 caacggaacc ccccacggaa cccccgcgac acccgctccc caattgcgtt agaacagcgg
      181 tggattgtcg gcttcgttgt gggccttttg agccgcttcc tgttctgccg cacgctcttt
      241 cctcgcccga tagccgagtc gcttaacggt gtccagatgc agcccgaaat gtttggccgt
      301 ttgcggccaa gagtggccct cgtcgtcgtg ataggcgcgg atgcgttcgc ggcgtgcagc
      361 ctgctcggcg agccactcgc tgcgttcctg cgccacgagc cggacgacgt ggcgttcgga
      421 tagtccggtg attcgagcgc cttcggcggc ggtcacgcgc cgctttttgc ggacagtcgg
      481 ctgccggttg tagccgtcgc tgtagccgtc gctcatagca atgcctccat ggctgacgcg
      541 gactttgcgc gccgcgcaac tgtgctcgcc gccgtgcgcg ctgctgcgcc cttccgcgag
      601 atggccgact ggcgcgcact gagtgtggcc tcgtagacca cgatcccgtc cgcccaaatg
      661 cgcgacttgg ttgtgatcca acgccaaatg ctgttggcga tggcgcggac ctcgctgtcc
      721 ggtagcggtc cgggacacac gtcgttgcac gggaactcgg cgtttcgcgc gtggcactcg
      781 gcatagatcg cgcggccgag tccgtccacg ttccgggtcg gcaggtagat ccgcatgagg
      841 gcgggacgat aggcccacaa cctgacggaa tcgaacagtg cgcaattccg ccctagcggc
      901 gtcggagccg ctttgtacgt ggtctgctga cgccagcgcg gcggtggcat gttcgcgccg
      961 agctcggcct cgatgtggct gagtgtgtag agatctgagt ggagccattc cgtttcccag
     1021 gcgatgtggc cggggttttt ggtcatgagg cctgagtaac tgcggtcgcc gtccacggcg
     1081 cgccgaaggc cttcggcgca cgccgccatg tatgcgagcg gcttacgccg cgcgtattcg
     1141 gtgcgtggaa caggggcgtt gagtgcccac actgcgtgtg cgtggccgtt ggcgcgattg
     1201 cccacgatcg cgttgggcag cggatgggac ccccgggcgc tgagcgctcg gagcgctgcg
     1261 tctggatggt ctacgtccac gaccagcagg tttgccagcg ctgttgggtt cgcctcgatg
     1321 taccggcggc ctagggccga cgcgcggctt tggcggtaga tcccctcgag cagatcgtcg
     1381 cttgccagcg gccagtacgg cagccagagc tgctcaaatt cgtcggcgac gtggctcacg
     1441 cttggtagta gaccacgatt aatcaccggt gtatggtccg acacgagctc caagtcagat
     1501 atttcgctga ggggccaccc cacaactgca cactcccccg ctctcccgtc gagccctggt
     1561 ggtggaacac cagcgacagc cgagcacccc caaccacctg taccaaccag gaggaacaca
     1621 tgcgtcgttt cgaggacgtt tccgggccgc tgagagccgc tgtggcggcc gtacacgccg
     1681 ccttagaccc gttagacccc ctgccgcctg aatgcgcggg tacgagccac acagcgcccg
     1741 aacttacgga gctggtgggc tcacctggct ttatggcgta cgaatcggct gtgtgcgacc
     1801 tgttgggcga ggtgaggtac gcgctactca cgctcgcttg agaccacccc aggatacata
     1861 gattaccaca actccgagcc cttccaccta gcgtgctaac cacgcacgct attgttgtat
     1921 cccatataat gcaatcgtag tctacctgac tgtacttaga aatgtggctt cgcctttgcc
     1981 cacgcacctg atcgctcctc gtttgctttt aaggaccgga cgaaccacag agcattagaa
     2041 ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat
     2101 agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc
     2161 cgacaatctc caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa
     2221 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2281 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2341 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2401 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2461 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2521 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2581 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2641 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2701 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2761 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2821 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2881 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
     2941 gcgtaaatgg gtagggggct tcaaatcgtc cccccatacg atataagttg ttactagtgc
     3001 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     3061 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
     3121 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     3181 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     3241 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     3301 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     3361 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     3421 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     3481 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     3541 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
     3601 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     3661 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     3721 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     3781 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     3841 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttgatacctg
     3901 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     3961 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     4021 ctggaataag gaagtgccat tccgcctgac ct
//
