LOCUS       sequence-401271-        3016 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 3016)
  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 3016)
  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_60 [sfGFP]
FEATURES             Location/Qualifiers
     source          1..3016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..795
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1060..1076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1060..1076)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1073..1095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1248..1836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1328..1347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1969..2063)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2084..2743)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2658..2677
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2744..2846)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2933..2955
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2947..2964
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2948..2964
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgcgtaaa ggcgaagaac tgttcactgg tgtcgtccct
      121 attctggtgg aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt
      181 gaaggtgacg caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg
      241 ccggtacctt ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt
      301 tatccggacc atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg
      361 caggaacgca cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa
      421 tttgaaggcg ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagat
      481 ggcaatatcc tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc
      541 gccgataaac aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat
      601 ggcagcgtgc agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt
      661 ctgctgccag acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag
      721 aaacgcgatc atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg
      781 gatgaactgt acaaatgagc tttgagacca ccccaggata catagattac cacaactccg
      841 agcccttcca cctagcgtgc taaccacgca cgctattgtt gtatcccata taatgcaatc
      901 gtagtctacc tgactgtact tagaaatgtg gcttcgcctt tgcccacgca cctgatcgct
      961 cctcgtttgc ttttaaggac cggacgaacc acagagcatt agaaggctag gtggaggctc
     1021 agtgatgata agtctgcgat ggtggatgca tgtgtcatgg tcatagctgt ttcctgtgtg
     1081 aaattgttat ccgctcagag ggcacaatcc tattccgcgc tatccgacaa tctccaagac
     1141 attaggtgga gttcagttcg gcgtatggca tatgtcgctg gaaagaacat gtgagcaaaa
     1201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1681 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1861 gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa gtcagcgtaa atgggtaggg
     1921 ggcttcaaat cgtcccccca tacgatataa gttgttacta gtgcttggat tctcaccaat
     1981 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     2041 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     2101 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     2161 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     2221 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     2281 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     2341 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     2401 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     2461 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     2521 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     2581 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     2641 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     2701 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     2761 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     2821 agttggaacc tcttacgtgc cgatcaacgt ctcattgata cctgaaacaa aacccatcgt
     2881 acggccaagg aagtctccaa taactgtgat ccaccacaag cgccagggtt ttcccagtca
     2941 cgacgttgta aaacgacggc cagtcatgca taatccgcac gcatctggaa taaggaagtg
     3001 ccattccgcc tgacct
//
