LOCUS       sequence-401233-        3545 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 3545)
  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 3545)
  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_16 [2u ORI]
FEATURES             Location/Qualifiers
     source          1..3545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(281..375)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(396..1055)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     970..989
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1056..1158)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1245..1267
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1259..1276
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1260..1276
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1413..2755
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    1745..1792
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(2917..2933)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2917..2933)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2930..2952)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(3105..3545,1..148))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3185..3204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
       61 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
      121 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      181 cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt aaatgggtag ggggcttcaa
      241 atcgtccccc catacgatat aagttgttac tagtgcttgg attctcacca ataaaaaacg
      301 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
      361 ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg
      421 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
      481 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
      541 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
      601 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
      661 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
      721 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
      781 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
      841 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
      901 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
      961 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     1021 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     1081 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     1141 cctcttacgt gccgatcaac gtctcattga tacctgaaac aaaacccatc gtacggccaa
     1201 ggaagtctcc aataactgtg atccaccaca agcgccaggg ttttcccagt cacgacgttg
     1261 taaaacgacg gccagtcatg cataatccgc acgcatctgg aataaggaag tgccattccg
     1321 cctgacctta gcgtgcgaac agcacgctat tgttgtatca ttactcgcat ccattctcag
     1381 gctgtctcgt ctcgtctccg aggtctccgg aggatccaat atcaaaggaa atgatagcat
     1441 tgaaggatga gactaatcca attgaggagt ggcagcatat agaacagcta aagggtagtg
     1501 ctgaaggaag catacgatac cccgcatgga atgggataat atcacaggag gtactagact
     1561 acctttcatc ctacataaat agacgcatat aagtacgcat ttaagcataa acacgcacta
     1621 tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
     1681 cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
     1741 ctttgaagtt cctattccga agttcctatt ctctagaaag tataggaact tcagagcgct
     1801 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     1861 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     1921 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     1981 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     2041 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     2101 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     2161 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     2221 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     2281 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     2341 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     2401 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     2461 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     2521 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     2581 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     2641 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     2701 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttcaggt
     2761 ggcacttttc ggggcgctcg agaccacccc aggatacata gattaccaca actccgagcc
     2821 cttccaccta gcgtgctaac cacgcacgct attgttgtat aggctaggtg gaggctcagt
     2881 gatgataagt ctgcgatggt ggatgcatgt gtcatggtca tagctgtttc ctgtgtgaaa
     2941 ttgttatccg ctcagagggc acaatcctat tccgcgctat ccgacaatct ccaagacatt
     3001 aggtggagtt cagttcggcg tatggcatat gtcgctggaa agaacatgtg agcaaaaggc
     3061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3541 agaag
//
