LOCUS       sequence-397987-        5044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 plasmid, barcoded ORI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5044)
  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 5044)
  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: pGL1_43
FEATURES             Location/Qualifiers
     source          1..5044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(144..160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(157..179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(332..920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(412..431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1056..1871)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1056..1871)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1779..1798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1863..1882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1992..2008
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1992..2008
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2009..2055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2169..2191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2183..2200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2184..2200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2937..4379
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGQHPGDCLRVDTAVA"
ORIGIN
        1 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
       61 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
      121 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      181 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
      241 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
      301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
      961 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     1021 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     1081 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1141 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1201 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1261 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1321 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1381 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1441 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1501 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1561 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1621 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1681 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1741 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1801 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     1861 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     1921 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     1981 tcccccgccg ctctagaact agtggatcca aataaaacga aaggctcagt cgaaagactg
     2041 ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga cgtccaggtt gggatacctg
     2101 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     2161 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     2221 ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg ctattgttgt
     2281 atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgagctct tcaatgagtc
     2341 agggtcacgc gtaggacgcg tcgagtaggg ttccgtttgt tcaaaggtca cgttccgcag
     2401 ccacacgccg tgtgaagctg gggaggatca acaggaccca gaaatggcac gagcccggga
     2461 ttggcctccc gggctcgccg cccccgaacc ggcgcagcat gccgagtgag gacgtttgct
     2521 ctgtgacgag cgtacgagag aacagcgcgt cgacaccacc gcgccccgcc ggaccgaagt
     2581 gccactgcgg caccccgatc gtccacatcc ccgggaagcg ccccaaaatc tactgctcgg
     2641 acgcctgccg aaagcgcgca aagcgtctca ttgcggacca gtcacgcagc gtagcaagca
     2701 cgactacagc gggtaatccg gacgcgggct ttagtagtaa gggaaaatct ccgtccactg
     2761 caggtcagag cgctgtaact ggaacgggtt ctaccccaaa cgagggtgcc gcgacagccg
     2821 ggcgggagga cccgcgcgac gagcggttcg cccggcggga ccggcaccag acggtgtcgc
     2881 tgaacgaggc gttcaagggc tgcgggaccc ggctcactgc cgggacggcc gagctgatgt
     2941 ggaagccggg cgaggcgacg tggggcaaca cgtgccgctg caacaacgtc cacacctgcc
     3001 cgtggtgcat gagccgcatc ctggccgtac ggggctcgaa cgtgcagctc gccgccgacg
     3061 gcctcgcgga cgccgggtac gggctgcacc ttggcacgaa cacgttgcgc cacttcgagc
     3121 gcatggcgtt cggcaccgtc cgcaagggaa tgcggcacgg cctggtcgcg gtcttgcacg
     3181 acggctggaa gggcgcgtac ggctcgtccg gccgccgctg gcgcacgatg cgcgacgact
     3241 tcgggatcat cggctacgag cgggccttcg aggacacctt cggctggggc tccggctggc
     3301 acctgcactg gcacacgctc tgggtcaccc gcgaggtgct cggcccggac gcccaggccg
     3361 cgttccgcga cgcactcgcc ggagcctggg ccgccggggt cgaatccgcg ggcggctaca
     3421 ccgtcagcga gacgtgcgac cggcccggtt gctcctgtga gggcaagggc cacggcaccg
     3481 acgtgcgccc actcaacggc gcggacgccg ctgacggcga tgcgggcaag caggcccgct
     3541 acctctacaa ggacggcgac aagaccaagg gcggcgtcgc caagatcggc ctggagctgg
     3601 ccggacagaa cttcaaggcc ggtcgcggcg acgaccgcat gggaccgctc gacctgggcg
     3661 acgcggcggc cgccgagctg cagcggcttc gtcggcccgg cccgttcgtc gagaagtacc
     3721 gcgagcgcga gttcggcgtc ttccaggtcc gcaagcacta ccggtcgcag aacctgaacc
     3781 ggctgatcaa ggaactcggc atccagcagg acgtccgcac cgaggaggag atcaccgacg
     3841 acaccgaggg cctggtcgcc atcgcggtca tcccggccta catctggtac cgatacatcg
     3901 cccgcgtcgc cggacgtcgc ctcgacctca tcaaggtcgc cgagacgtac ggcctaccgg
     3961 gcgtccggcg gctcgtcgag tcctggggcc tggtgtgggg caaggacgtc ctggacccgc
     4021 ccgccccgga agcgccggcc gcgccgggcg accttgacgc tgaccagatg aggttcgagg
     4081 tcatgtccga ggaggaagct gctttccgcg aggcgcgccg gaaggccaac gaggcccgca
     4141 cggaggagct ggccgcttcg ctcgaccgag tgcgtcagcc gaagaaggag gcgatccgcc
     4201 cgacgatcag ccttcggaag cgcctgaagc cgaagccggt caccgtcgac gtcaagactc
     4261 ccccgcccgg cgccgcgtcg cccgtgtgcc gccgctgcaa gggcaagctc gccccggtac
     4321 tccagccctg gggccagcac cccggagact gcctccgcgt cgacacagcg gtcgcgtagg
     4381 gccgctgagg gaccgccatc agaccgccac cccggacatc cgggtggcgg tctgatggcg
     4441 gtcccggcgc gaaagccgag ggtctgcctg ccgtgaggtg gccggcggac ctgggttact
     4501 gctggaatcc cgcgcccagt ccgagcgtcc gccggcgctt ttcacctggc tccgggtgct
     4561 cgtcgatgag caccaggccg tcgacgacga cgcgtccgcc tgcctgcttt tggccggcgg
     4621 gccgggccgc ggttgtccgc cgggtcggac cggtcccttg gtcgaccgat gcctcggcca
     4681 cgacgatgct gggcccgaag cggtacgcga cccgtgccgg gccgcagcgg atcgcggcag
     4741 caggagctcc ccgcccgcca cggctggctg gatggtcgcc tcacgcgccc gcaggggtga
     4801 cggcgtgggt gcggcggtca gccccgtgat cgggcgtcag acggccgctg agggaccgcg
     4861 accacccctc ccttcccgcc tgccgtcttt caccgcccgg ccggtccgca gacgccgtac
     4921 ggccaggcca cggaggatta cgggcagatg gcgcccaccc ggcacccggg ctggggcacc
     4981 ctggacagca cgaaggtccg ccccactcac cggggcgggc cttcgcgttg agctagagtt
     5041 cgct
//
