LOCUS       sequence-402923-        2959 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 2959)
  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 2959)
  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_33 [cat]
FEATURES             Location/Qualifiers
     source          1..2959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..741
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1003..1019)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1003..1019)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1016..1038)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1191..1779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1271..1290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1912..2006)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2027..2686)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(2687..2789)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2876..2898
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2890..2907
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2891..2907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatggagaaa aaaatcactg gatataccac cgttgatata
      121 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
      181 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
      241 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
      301 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
      361 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      421 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      481 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      541 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      601 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      661 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      721 gagtggcagg gcggggcgta agctttgaga ccaccccagg atacatagat taccacaact
      781 ccgagccctt ccacctagcg tgctaaccac gcacgctatt gttgtatccc atataatgca
      841 atcgtagtct acctgactgt acttagaaat gtggcttcgc ctttgcccac gcacctgatc
      901 gctcctcgtt tgcttttaag gaccggacga accacagagc attagaaggc taggtggagg
      961 ctcagtgatg ataagtctgc gatggtggat gcatgtgtca tggtcatagc tgtttcctgt
     1021 gtgaaattgt tatccgctca gagggcacaa tcctattccg cgctatccga caatctccaa
     1081 gacattaggt ggagttcagt tcggcgtatg gcatatgtcg ctggaaagaa catgtgagca
     1141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1801 acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg taaatgggta
     1861 gggggcttca aatcgtcccc ccatacgata taagttgtta ctagtgcttg gattctcacc
     1921 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1981 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
     2041 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     2101 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     2161 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     2221 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     2281 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     2341 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     2401 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2461 ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2521 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2581 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2641 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2701 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2761 gaaagttgga acctcttacg tgccgatcaa cgtctcattg atacctgaaa caaaacccat
     2821 cgtacggcca aggaagtctc caataactgt gatccaccac aagcgccagg gttttcccag
     2881 tcacgacgtt gtaaaacgac ggccagtcat gcataatccg cacgcatctg gaataaggaa
     2941 gtgccattcc gcctgacct
//
