LOCUS       sequence-397981-        2915 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 2915)
  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 2915)
  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_114 [p15A]
FEATURES             Location/Qualifiers
     source          1..2915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(356..450)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(471..1130)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1045..1064
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1131..1233)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1320..1342
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1334..1351
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1335..1351
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1524..2069
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2186..2203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2362..2378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2362..2378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2375..2397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(2550..2915,1..223))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
       61 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
      121 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
      181 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
      241 ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc agcgtaaatg
      301 ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg cttggattct
      361 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
      421 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg
      481 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
      541 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
      601 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
      661 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
      721 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
      781 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
      841 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
      901 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
      961 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     1021 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     1081 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     1141 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     1201 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct gaaacaaaac
     1261 ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc cagggttttc
     1321 ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca tctggaataa
     1381 ggaagtgcca ttccgcctga ccttagcgtg cgaacagcac gctattgttg tatcattact
     1441 cgcatccatt ctcaggctgt ctcgtctcgt ctccgaggtc tccggagcta gaaatatttt
     1501 atctgattaa taagatgatc ttcttgagat cgttttggtc tgcgcgtaat ctcttgctct
     1561 gaaaacgaaa aaaccgcctt gcagggcggt ttttcgaagg ttctctgagc taccaactct
     1621 ttgaaccgag gtaactggct tggaggagcg cagtcaccaa aacttgtcct ttcagtttag
     1681 ccttaaccgg cgcatgactt caagactaac tcctctaaat caattaccag tggctgctgc
     1741 cagtggtgct tttgcatgtc tttccgggtt ggactcaaga cgatagttac cggataaggc
     1801 gcagcggtcg gactgaacgg ggggttcgtg catacagtcc agcttggagc gaactgccta
     1861 cccggaactg agtgtcaggc gtggaatgag acaaacgcgg ccataacagc ggaatgacac
     1921 cggtaaaccg aaaggcagga acaggagagc gcacgaggga gccgccaggg ggaaacgcct
     1981 ggtatcttta tagtcctgtc gggtttcgcc accactgatt tgagcgtcag atttcgtgat
     2041 gcttgtcagg ggggcggagc ctatggaaaa acggctttgc cgcggccctc tcacttccct
     2101 gttaagtatc ttcctggcat cttccaggaa atctccgccc cgttcgtaag ccatttccgc
     2161 tcgccgcagt cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aatatatccc
     2221 gctcgagacc accccaggat acatagatta ccacaactcc gagcccttcc acctagcgtg
     2281 ctaaccacgc acgctattgt tgtataggct aggtggaggc tcagtgatga taagtctgcg
     2341 atggtggatg catgtgtcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcag
     2401 agggcacaat cctattccgc gctatccgac aatctccaag acattaggtg gagttcagtt
     2461 cggcgtatgg catatgtcgc tggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2881 gacttatcgc cactggcagc agccactggt aacag
//
