LOCUS       sequence-408502-        4358 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 4358)
  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 4358)
  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_4 [pIJ101 ORI]
FEATURES             Location/Qualifiers
     source          1..4358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(330..424)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(445..1104)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1019..1038
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1105..1207)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1294..1316
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1308..1325
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1309..1325
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3779..3795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3779..3795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3792..3814)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(3967..4358,1..197))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4047..4066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
       61 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
      121 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
      181 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctctattca
      241 acaaagccgc cgtcccgtca agtcagcgta aatgggtagg gggcttcaaa tcgtcccccc
      301 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
      361 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      421 agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg
      481 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
      541 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
      601 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
      661 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
      721 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
      781 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
      841 aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt ccggatgagc
      901 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
      961 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
     1021 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
     1081 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
     1141 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     1201 ccgatcaacg tctcattgat acctgaaaca aaacccatcg tacggccaag gaagtctcca
     1261 ataactgtga tccaccacaa gcgccagggt tttcccagtc acgacgttgt aaaacgacgg
     1321 ccagtcatgc ataatccgca cgcatctgga ataaggaagt gccattccgc ctgaccttag
     1381 cgtgcgaaca gcacgctatt gttgtatcat tactcgcatc cattctcagg ctgtctcgtc
     1441 tcgtctccga ggtctccgga gccatcgctc gccgacacac gcactgaccg cacgtcaaag
     1501 ccccgccgga tcaccggcgg ggctctcttc ggccctccaa gtcacaccag ccccaagggg
     1561 cgtcgggagt ggcggaggga acctctggcc cgattggtgc caggattccc accagaccaa
     1621 agagcaacgg gccggacttc gcacctccga cccgtccgct cccagactcg cgccccttag
     1681 ccgggcgaga caggaacgtt gctcgtgccc agagtacgga gcgatgccga ggcattgcca
     1741 gatcggcccg ccgggccccg ctgccactgc gggaccgcaa ttgcccacac accgggcaaa
     1801 cggccgcgta tctactgctc agaccgctgc cggatggcag cgaagcgggc gatcgcgcgt
     1861 gtgacgcgag atgccgcccg aggcaaaagc gaacaccttg ggaaagaaac aacagagttt
     1921 cccgcacccc tccgacctgc ggtttctccg gacggggtgg atggggagag cccgagaggc
     1981 gacagcctct cggaagtagg aagcacgtcg cggagcgacg ctgcccgact gcggaaagcc
     2041 gcccggtaca gccgccgccg gacgctgtgg cggatcagcg gggacgccgc gtgcaagggc
     2101 tgcggccgcg ccctgatgga ccctgcctcc ggcgtaatcg tcgcccagac ggcggccgga
     2161 acgtccgtgg tcctgggcct gatgcggtgc gggcggatct ggctctgccc ggtctgcgcc
     2221 gccacgatcc ggcacaagcg ggccgaggag atcaccgccg ccgtggtcga gtggatcaag
     2281 cgcgggggga ccgcctacct ggtcaccttc acggcccgcc atgggcacac ggaccggctc
     2341 gcggacctca tggacgccct ccagggcacc cggaagacgc cggacagccc ccggcggccg
     2401 ggcgcctacc agcgactgat cacgggcggc acgtgggccg gacgccgggc caaggacggg
     2461 caccgggccg ccgaccgcga gggcatccga gatcggatcg ggtacgtcgg catgatccgc
     2521 gcgaccgaag tcaccgtggg gcagatcaac ggctggcacc cgcacatcca cgcgatcgtc
     2581 ctggtcggcg gccggaccga gggggagcgg tccgcgaagc agatcgtcgc caccttcgag
     2641 ccgaccggcg ccgcgctcga cgagtggcag gggcactggc ggtccgtgtg gaccgccgcc
     2701 ctgcgcaagg tcaaccccgc cttcacgccc gacgaccggc acggcgtcga cttcaagcgg
     2761 ctggaaaccg agcgcgacgc caacgacctc gccgagtaca tcgccaagac ccaggacggg
     2821 aaggcgcccg ccctcgaact cgcccgcgcc gacctcaaga cggcgaccgg cgggaacgtc
     2881 gccccgttcg aactcctcgg acggatcggg gacctgaccg gcggcatgac cgaggacgac
     2941 gccgccgggg tcggctcgct ggagtggaac ctctcgcgct ggcacgagta cgagcgggca
     3001 acccggggac gccgggccat cgaatggacc cgctacctgc ggcagatgct cgggctcgac
     3061 ggcggcgaca ccgaggccga cgacctcgat ctgctcctgg cggccgacgc cgacggcggg
     3121 gagctgcggg ccggggtcgc cgtgaccgag gacggatggc acgcggtcac ccgccgcgcc
     3181 ctcgacctcg aggcgacccg ggccgccgaa ggcaaggacg gcaacgagga ttcggcggcc
     3241 gtgggcgaac gggtgcggga ggtcctggcg ctggccgacg cggccgacac agtggtggtg
     3301 ctcacggcgg gggaggtggc cgaggcgtac gccgacatgc tcgccgccct cgcccagcgc
     3361 cgcgaggaag caactgcacg ccgacggcga gagcaggacg acgaccagga cgacgacgcc
     3421 gacgaccgcc aggagcgggc cgcccggcac atcgcccggc tcgcaagtgg gcccacttcg
     3481 cactaactcg ctcccccccg ccgtacgtca tcccggtgac gtacggcggg ggtcggtgac
     3541 gtacgcggcg acggcggccg gggtcgaagc cgcgggagta atcctgggat tactcgcccg
     3601 gggtcggccc cgccggcact tcgtgcaggc ggtacccgct cgagaccacc ccaggataca
     3661 tagattacca caactccgag cccttccacc tagcgtgcta accacgcacg ctattgttgt
     3721 ataggctagg tggaggctca gtgatgataa gtctgcgatg gtggatgcat gtgtcatggt
     3781 catagctgtt tcctgtgtga aattgttatc cgctcagagg gcacaatcct attccgcgct
     3841 atccgacaat ctccaagaca ttaggtggag ttcagttcgg cgtatggcat atgtcgctgg
     3901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4321 cactggtaac aggattagca gagcgaggta tgtaggcg
//
