LOCUS       sequence-401274-        2422 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 2422)
  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 2422)
  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_32 [Pcat]
FEATURES             Location/Qualifiers
     source          1..2422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        102..204
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(466..482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(466..482)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(479..501)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(654..1242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(734..753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1375..1469)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1490..2149)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2064..2083
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2150..2252)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2339..2361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2353..2370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2354..2370
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagtggcga aaatgagacg ttgatcggca cgtaagaggt
      121 tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga
      181 ttttcaggag ctaaggaagc taaaaatgtg agaccacccc aggatacata gattaccaca
      241 actccgagcc cttccaccta gcgtgctaac cacgcacgct attgttgtat cccatataat
      301 gcaatcgtag tctacctgac tgtacttaga aatgtggctt cgcctttgcc cacgcacctg
      361 atcgctcctc gtttgctttt aaggaccgga cgaaccacag agcattagaa ggctaggtgg
      421 aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc
      481 tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc cgacaatctc
      541 caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa gaacatgtga
      601 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      661 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      721 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      781 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      841 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      901 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      961 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1021 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1081 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1141 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1201 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1261 tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca gcgtaaatgg
     1321 gtagggggct tcaaatcgtc cccccatacg atataagttg ttactagtgc ttggattctc
     1381 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     1441 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     1501 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1561 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1621 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     1681 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     1741 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
     1801 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
     1861 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
     1921 ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
     1981 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
     2041 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
     2101 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
     2161 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     2221 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttgatacctg aaacaaaacc
     2281 catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc agggttttcc
     2341 cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat ctggaataag
     2401 gaagtgccat tccgcctgac ct
//
