LOCUS       sequence-401250-        3160 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 3160)
  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 3160)
  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_26 [bla pBR322]
FEATURES             Location/Qualifiers
     source          1..3160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(82..150,151..942)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1204..1220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1204..1220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1217..1239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1392..1980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1472..1491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2113..2207)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2228..2887)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2802..2821
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2888..2990)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3077..3099
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3091..3108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3092..3108
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgagcatt cagcattttc gtgtggcgct gattccgttt
      121 tttgcggcgt tttgcctgcc ggtgtttgcg catccggaaa ccctggtgaa agtgaaagat
      181 gcggaagatc aactgggtgc gcgcgtgggc tatattgaac tggatctgaa cagcggcaaa
      241 attctggaat cttttcgtcc ggaagaacgt tttccgatga tgagcacctt taaagtgctg
      301 ctgtgcggtg cggttctgag ccgtgtggat gcgggccagg aacaactggg ccgtcgtatt
      361 cattatagcc agaacgatct ggtggaatat agcccggtga ccgaaaaaca tctgaccgat
      421 ggcatgaccg tgcgtgaact gtgcagcgcg gcgattacca tgagcgataa caccgcggcg
      481 aacctgctgc tgacgaccat tggcggtccg aaagaactga ccgcgtttct gcataacatg
      541 ggcgatcatg tgacccgtct ggatcgttgg gaaccggaac tgaacgaagc gattccgaac
      601 gatgaacgtg ataccaccat gccggcagca atggcgacca ccctgcgtaa actgctgacg
      661 ggtgagctgc tgaccctggc aagccgccag caactgattg attggatgga agcggataaa
      721 gtggcgggtc cgctgctgcg tagcgcgctg ccggctggct ggtttattgc ggataaaagc
      781 ggtgcgggcg aacgtggcag ccgtggcatt attgcggcgc tgggcccgga tggtaaaccg
      841 agccgtattg tggtgattta taccaccggc agccaggcga cgatggatga acgtaaccgt
      901 cagattgcgg aaattggcgc gagcctgatt aaacattggt aagctttgag accaccccag
      961 gatacataga ttaccacaac tccgagccct tccacctagc gtgctaacca cgcacgctat
     1021 tgttgtatcc catataatgc aatcgtagtc tacctgactg tacttagaaa tgtggcttcg
     1081 cctttgccca cgcacctgat cgctcctcgt ttgcttttaa ggaccggacg aaccacagag
     1141 cattagaagg ctaggtggag gctcagtgat gataagtctg cgatggtgga tgcatgtgtc
     1201 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc agagggcaca atcctattcc
     1261 gcgctatccg acaatctcca agacattagg tggagttcag ttcggcgtat ggcatatgtc
     1321 gctggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1981 gaagatcctt tgatcttttc tacggggtct gacgctctat tcaacaaagc cgccgtcccg
     2041 tcaagtcagc gtaaatgggt agggggcttc aaatcgtccc cccatacgat ataagttgtt
     2101 actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa
     2161 atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga
     2221 tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     2281 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     2341 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     2401 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     2461 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     2521 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     2581 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     2641 ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa
     2701 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     2761 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     2821 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     2881 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     2941 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     3001 gatacctgaa acaaaaccca tcgtacggcc aaggaagtct ccaataactg tgatccacca
     3061 caagcgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtca tgcataatcc
     3121 gcacgcatct ggaataagga agtgccattc cgcctgacct
//
