LOCUS       sequence-401221-        2436 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 2436)
  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 2436)
  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_25 [P3 promoter]
FEATURES             Location/Qualifiers
     source          1..2436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        127..218
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(480..496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(480..496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(493..515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(668..1256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(748..767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1389..1483)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1504..2163)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2078..2097
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2164..2266)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2353..2375
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2367..2384
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2368..2384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagtagtag cccgcctaat gagcgggctt ttttttaatt
      121 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      181 cctgataaat gcttcaataa tattgaaaaa ggaagagtaa tgtgagacca ccccaggata
      241 catagattac cacaactccg agcccttcca cctagcgtgc taaccacgca cgctattgtt
      301 gtatcccata taatgcaatc gtagtctacc tgactgtact tagaaatgtg gcttcgcctt
      361 tgcccacgca cctgatcgct cctcgtttgc ttttaaggac cggacgaacc acagagcatt
      421 agaaggctag gtggaggctc agtgatgata agtctgcgat ggtggatgca tgtgtcatgg
      481 tcatagctgt ttcctgtgtg aaattgttat ccgctcagag ggcacaatcc tattccgcgc
      541 tatccgacaa tctccaagac attaggtgga gttcagttcg gcgtatggca tatgtcgctg
      601 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      661 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      721 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      781 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      841 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      901 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      961 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1021 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1081 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1141 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1201 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1261 atcctttgat cttttctacg gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa
     1321 gtcagcgtaa atgggtaggg ggcttcaaat cgtcccccca tacgatataa gttgttacta
     1381 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     1441 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc
     1501 aaattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     1561 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     1621 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     1681 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     1741 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     1801 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     1861 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     1921 ctcaccgtct ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat
     1981 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     2041 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     2101 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     2161 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     2221 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattgata
     2281 cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag
     2341 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca taatccgcac
     2401 gcatctggaa taaggaagtg ccattccgcc tgacct
//
