LOCUS       sequence-397980-        3218 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 3218)
  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 3218)
  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_142 [EilR-PjexD]
FEATURES             Location/Qualifiers
     source          1..3218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             799..807
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(1262..1278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1262..1278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1275..1297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1450..2038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1530..1549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2171..2265)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2286..2945)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2860..2879
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2946..3048)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3135..3157
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3149..3166
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3150..3166
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagggcccg acgcgcgccc agtcgacggg gaatttgccg
      121 agatggggga ccggcgcgag cgcgtagaag gtcgaatggc ccggcggcgc catttttggg
      181 tcggtcgcgg tcgggtgatg gagatagagc gagttacgaa aataactcaa gctgaataac
      241 gtgctgcagg tggcgggtga atgcggcatc gtctacctcc ggcattccca gcacgtaaat
      301 cccgtccagt ccacagacca gcgaaatcag ccgccaggcg atattttcgg cgctatcgcg
      361 cagggtaaat tcgccagcgg catggcccgc gcgaatgatc ctgaccgctt cgtcatgcca
      421 caggttcatg gtcagcaggt acgcgctttt gatttccgga tcgctgtcgg ccagcagctg
      481 cgcctggcgc cacaggcgga tatagggctc cagccgtccg tcctcgctgc cgagggcgga
      541 aaacagctgt tcacgccagc cggcggtgcg cgaaaggcgc tgcagatcca tcatctcgcg
      601 gatcacgcgg ataaacgcct gggattttag ctcgccggag gaggtgaaat ggtgatggac
      661 ctggcctgcg gcaacacccg ctgcggtggc gatgttgcgg accgtcatac cggtaaaccc
      721 ttgatccagc gcgacgcgca tggcggcctg catgatggtt tcccgacgtt cttcgcgatt
      781 cagatagccc atattgtcac ctccttatcc acacattata cgagccggat gattaatggg
      841 gaaccttacc agagggcgcc ccagctggca attcctctta agtaagtaag agtatacgta
      901 tatcggctaa taacgtatta aggcgcttcg gcgccttttt ttatgggggt attttcatcc
      961 caatccacac gtccaacgca cagcaaacac cacgtcgacc ctatcagctg cgtgctttct
     1021 atgagtcgtt gctgcataac aaaaaactgc aaaaaatagt ttgacaggac acgtgtccaa
     1081 ctttaagatg tacccaaagt tggacacgtg tccaactttt acttgagacc accccaggat
     1141 acatagatta ccacaactcc gagcccttcc acctagcgtg ctaaccacgc acgctattgt
     1201 tgtataggct aggtggaggc tcagtgatga taagtctgcg atggtggatg catgtgtcat
     1261 ggtcatagct gtttcctgtg tgaaattgtt atccgctcag agggcacaat cctattccgc
     1321 gctatccgac aatctccaag acattaggtg gagttcagtt cggcgtatgg catatgtcgc
     1381 tggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2041 agatcctttg atcttttcta cggggtctga cgctctattc aacaaagccg ccgtcccgtc
     2101 aagtcagcgt aaatgggtag ggggcttcaa atcgtccccc catacgatat aagttgttac
     2161 tagtgcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     2221 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     2281 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     2341 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     2401 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     2461 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     2521 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     2581 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     2641 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     2701 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     2761 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     2821 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     2881 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     2941 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     3001 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcattga
     3061 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     3121 agcgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc
     3181 acgcatctgg aataaggaag tgccattccg cctgacct
//
