LOCUS       sequence-393056-        3822 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 3822)
  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 3822)
  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_9 [pUB110 ORI]
FEATURES             Location/Qualifiers
     source          1..3822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             556..1560
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     primer_bind     complement(1866..1882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1879..1901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2054..2642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2134..2153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2775..2869)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2890..3549)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3464..3483
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3550..3652)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3739..3761
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3753..3770
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3754..3770
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggagctaaaa taagcgattt aaagcgtgat gtgagtttaa
      121 tctatgaaag cactaaggaa ttccttaagg aacgtacaga cggcttaaaa gcctttaaaa
      181 acgtttttaa ggggtttgta gacaaggtaa aggataaaac agcacaattc caagaaaaac
      241 acgatttaga acctaaaaag aacgaatttg aactaactca taaccgagag gtaaaaaaag
      301 aacgaagtcg agatcaggga atgagtttat aaaataaaaa aagcacctga aaaggtgtct
      361 ttttttgatg gttttgaact tgttctttct tatcttgata catatagaaa taacgtcatt
      421 tttattttag ttgctgaaag gtgcgttgaa gtgttggtat gtatgtgttt taaagtattg
      481 aaaaccctta aaattggttg cacagaaaaa ccccatctgt taaagttata agtgactaaa
      541 caaataacta aatagatggg ggtttctttt aatattatgt gtcctaatag tagcatttat
      601 tcagatgaaa aatcaagggt tttagtggac aagacaaaaa gtggaaaagt tagaccatgg
      661 agagaaaaga aaatcgctaa tgttgattac tttgaacttc tgcatattct tgaatttaaa
      721 aaggctgaaa gagtaaaaga ttgtgctgaa atattagagt ataaacaaaa tcgtgaaaca
      781 ggcgaaagaa agttgtatcg agtgtggttt tgtaaatcca ggctttgtcc aatgtgcaac
      841 tggaggagag caatgaaaca tggcattcag tcacaaaagg ttgttgctga agttattaaa
      901 caaaagccaa cagttcgttg gttgtttctc acattaacag ttaaaaatgt ttatgatggc
      961 gaagaattaa ataagagttt gtcagatatg gctcaaggat ttcgccgaat gatgcaatat
     1021 aaaaaaatta ataaaaatct tgttggtttt atgcgtgcaa cggaagtgac aataaataat
     1081 aaagataatt cttataatca gcacatgcat gtattggtat gtgtggaacc aacttatttt
     1141 aagaatacag aaaactacgt gaatcaaaaa caatggattc aattttggaa aaaggcaatg
     1201 aaattagact atgatccaaa tgtaaaagtt caaatgattc gaccgaaaaa taaatataaa
     1261 tcggatatac aatcggcaat tgacgaaact gcaaaatatc ctgtaaagga tacggatttt
     1321 atgaccgatg atgaagaaaa gaatttgaaa cgtttgtctg atttggagga aggtttacac
     1381 cgtaaaaggt taatctccta tggtggtttg ttaaaagaaa tacataaaaa attaaacctt
     1441 gatgacacag aagaaggcga tttgattcat acagatgatg acgaaaaagc cgatgaagat
     1501 ggattttcta ttattgcaat gtggaattgg gaacggaaaa attattttat taaagagtag
     1561 ttcaacaaac gggccagttt gttgaagatt agatgctata attgcgcttg agaccacccc
     1621 aggatacata gattaccaca actccgagcc cttccaccta gcgtgctaac cacgcacgct
     1681 attgttgtat cccatataat gcaatcgtag tctacctgac tgtacttaga aatgtggctt
     1741 cgcctttgcc cacgcacctg atcgctcctc gtttgctttt aaggaccgga cgaaccacag
     1801 agcattagaa ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg
     1861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt
     1921 ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt atggcatatg
     1981 tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2461 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2641 aagaagatcc tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc
     2701 cgtcaagtca gcgtaaatgg gtagggggct tcaaatcgtc cccccatacg atataagttg
     2761 ttactagtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     2821 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     2881 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     2941 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     3001 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     3061 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     3121 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3181 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3241 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3301 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
     3361 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     3421 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     3481 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     3541 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     3601 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     3661 ttgatacctg aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac
     3721 cacaagcgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat
     3781 ccgcacgcat ctggaataag gaagtgccat tccgcctgac ct
//
