LOCUS       sequence-401279-        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_24 [pBBR1-UP ORI]
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(72..731)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     646..665
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(732..834)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     921..943
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     935..952
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     936..952
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1176..1947
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1948..2610
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     complement(2928..2944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2928..2944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2941..2963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3116..3704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3196..3215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(3837..3880,1..51))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
       61 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
      121 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
      181 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
      241 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
      301 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
      361 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
      421 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
      481 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
      541 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
      601 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
      661 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
      721 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
      781 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct
      841 cattgatacc tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata actgtgatcc
      901 accacaagcg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtcatgcata
      961 atccgcacgc atctggaata aggaagtgcc attccgcctg accttagcgt gcgaacagca
     1021 cgctattgtt gtatcattac tcgcatccat tctcaggctg tctcgtctcg tctccgaggt
     1081 ctcaggagcg ccctacgggc ttgctctccg ggcttcgccc tgcgcggtcg ctgcgctccc
     1141 ttgccagccc gtggatatgt ggacgatggc cgcgagcggc caccggctgg ctcgcttcgc
     1201 tcggcccgtg gacaaccctg ctggacaagc tgatggacag gctgcgcctg cccacgagct
     1261 tgaccacagg gattgcccac cggctaccca gccttcgacc acatacccac cggctccaac
     1321 tgcgcggcct gcggccttgc cccatcaatt tttttaattt tctctgggga aaagcctccg
     1381 gcctgcggcc tgcgcgcttc gcttgccggt tggacaccaa gtggaaggcg ggtcaaggct
     1441 cgcgcagcga ccgcgcagcg gcttggcctt gacgcgcctg gaacgaccca agcctatgcg
     1501 agtgggggca gtcgaagggc gaagcccgcc cgcctgcccc ccgagcctca cggcggcgag
     1561 tgcgggggtt ccaagggggc agcgccacct tgggcaaggc cgaaggccgc gcagtcgatc
     1621 aacaagcccc ggaggggcca ctttttgccg gagggggagc cgcgccgaag gcgtggggga
     1681 accccgcagg ggtgcccttc tttgggcacc aaagaactag atatagggcg aaatgcgaaa
     1741 gacttaaaaa tcaacaactt aaaaaagggg ggtacgcaac agctcattgc ggcacccccc
     1801 gcaatagctc attgcgtagg ttaaagaaaa tctgtaattg actgccactt ttacgcaacg
     1861 cataattgtt gtcgcgctgc cgaaaagttg cagctgattg cgcatggtgc cgcaaccgtg
     1921 cggcacccct accgcatgga gataagcatg gccacgcagt ccagagaaat cggcattcaa
     1981 gccaagaaca agcccggtca ctgggtgcaa acggaacgca aagcgcatga ggcgtgggcc
     2041 gggcttattg cgaggaaacc cacggcggca atgctgctgc atcacctcgt ggcgcagatg
     2101 ggccaccaga acgccgtggt ggtcagccag aagacacttt ccaagctcat cggacgttct
     2161 ttgcggacgg tccaatacgc agtcaaggac ttggtggccg agcgctggat ctccgtcgtg
     2221 aagctcaacg gccccggcac cgtgttggcc tacgtggtca atgaccgcgt ggcgtggggc
     2281 cagccccgcg accagttgcg cctgtcggtg ttcagtgccg ccgtggtggt tgatcacgac
     2341 gaccaggacg aatcgctgtt ggggcatggc gacctgcgcc gcatcccgac cctgtatccg
     2401 ggcgagcagc aactaccgac cggccccggc gaggagccgc ccagccagcc cggcattccg
     2461 ggcatggaac cagacctgcc agccttgacc gaaacggagg aatgggaacg gcgcgggcag
     2521 cagcgcctgc cgatgcccga tgagccgtgt tttctggacg atggcgagcc gttggagccg
     2581 ccgacacggg tcacgctgcc gcgccggtag cacttgggtt gcgcagcaac ccgtaagtgc
     2641 gctgttccag actatcggct gtagcccgct tgagaccacc ccaggataca tagattacca
     2701 caactccgag cccttccacc tagcgtgcta accacgcacg ctattgttgt atcccatata
     2761 atgcaatcgt agtctacctg actgtactta gaaatgtggc ttcgcctttg cccacgcacc
     2821 tgatcgctcc tcgtttgctt ttaaggaccg gacgaaccac agagcattag aaggctaggt
     2881 ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc atagctgttt
     2941 cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta tccgacaatc
     3001 tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga aagaacatgt
     3061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3721 tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat
     3781 gggtaggggg cttcaaatcg tccccccata cgatataagt tgttactagt gcttggattc
     3841 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
//
