LOCUS       sequence-401290-        3880 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 3880)
  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 3880)
  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_2 [pBBR1 ORI]
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      172..943
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             944..1606
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     complement(1924..1940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1924..1940)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1937..1959)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2112..2700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2192..2211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2833..2927)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2948..3607)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3522..3541
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3608..3710)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3797..3819
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3811..3828
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3812..3828
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagcgccct acgggcttgc tctccgggct tcgccctgcg
      121 cggtcgctgc gctcccttgc cagcccgtgg atatgtggac gatggccgcg agcggccacc
      181 ggctggctcg cttcgctcgg cccgtggaca accctgctgg acaagctgat ggacaggctg
      241 cgcctgccca cgagcttgac cacagggatt gcccaccggc tacccagcct tcgaccacat
      301 acccaccggc tccaactgcg cggcctgcgg ccttgcccca tcaatttttt taattttctc
      361 tggggaaaag cctccggcct gcggcctgcg cgcttcgctt gccggttgga caccaagtgg
      421 aaggcgggtc aaggctcgcg cagcgaccgc gcagcggctt ggccttgacg cgcctggaac
      481 gacccaagcc tatgcgagtg ggggcagtcg aagggcgaag cccgcccgcc tgccccccga
      541 gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg caaggccgaa
      601 ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg gggagccgcg
      661 ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag aactagatat
      721 agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta cgcaacagct
      781 cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg taattgactg
      841 ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc tgattgcgca
      901 tggtgccgca accgtgcggc acccctaccg catggagata agcatggcca cgcagtccag
      961 agaaatcggc attcaagcca agaacaagcc cggtcactgg gtgcaaacgg aacgcaaagc
     1021 gcatgaggcg tgggccgggc ttattgcgag gaaacccacg gcggcaatgc tgctgcatca
     1081 cctcgtggcg cagatgggcc accagaacgc cgtggtggtc agccagaaga cactttccaa
     1141 gctcatcgga cgttctttgc ggacggtcca atacgcagtc aaggacttgg tggccgagcg
     1201 ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg tggtcaatga
     1261 ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg tcggtgttca gtgccgccgt
     1321 ggtggttgat cacgacgacc aggacgaatc gctgttgggg catggcgacc tgcgccgcat
     1381 cccgaccctg tatccgggcg agcagcaact accgaccggc cccggcgagg agccgcccag
     1441 ccagcccggc attccgggca tggaaccaga cctgccagcc ttgaccgaaa cggaggaatg
     1501 ggaacggcgc gggcagcagc gcctgccgat gcccgatgag ccgtgttttc tggacgatgg
     1561 cgagccgttg gagccgccga cacgggtcac gctgccgcgc cggtagcact tgggttgcgc
     1621 agcaacccgt aagtgcgctg ttccagacta tcggctgtag cccgcttgag accaccccag
     1681 gatacataga ttaccacaac tccgagccct tccacctagc gtgctaacca cgcacgctat
     1741 tgttgtatcc catataatgc aatcgtagtc tacctgactg tacttagaaa tgtggcttcg
     1801 cctttgccca cgcacctgat cgctcctcgt ttgcttttaa ggaccggacg aaccacagag
     1861 cattagaagg ctaggtggag gctcagtgat gataagtctg cgatggtgga tgcatgtgtc
     1921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc agagggcaca atcctattcc
     1981 gcgctatccg acaatctcca agacattagg tggagttcag ttcggcgtat ggcatatgtc
     2041 gctggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2701 gaagatcctt tgatcttttc tacggggtct gacgctctat tcaacaaagc cgccgtcccg
     2761 tcaagtcagc gtaaatgggt agggggcttc aaatcgtccc cccatacgat ataagttgtt
     2821 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     2881 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     2941 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3001 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     3061 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     3121 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     3181 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     3241 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     3301 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     3361 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     3421 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     3481 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     3541 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     3601 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     3661 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     3721 gatacctgaa acaaaaccca tcgtacggcc aaggaagtct ccaataactg tgatccacca
     3781 caagcgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtca tgcataatcc
     3841 gcacgcatct ggaataagga agtgccattc cgcctgacct
//
