LOCUS       sequence-408500-        4604 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 4604)
  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 4604)
  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_38
FEATURES             Location/Qualifiers
     source          1..4604
                     /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"
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 ttgtcagggg tagtgtaaca cgttccgcag
     2401 ccacacgccg tgtgaagctg gggagccatc gctcgccgac acacgcactg accgcacgtc
     2461 aaagccccgc cggatcaccg gcggggctct cttcggccct ccaagtcaca ccagccccaa
     2521 ggggcgtcgg gagtggcgga gggaacctct ggcccgattg gtgccaggat tcccaccaga
     2581 ccaaagagca acgggccgga cttcgcacct ccgacccgtc cgctcccaga ctcgcgcccc
     2641 ttagccgggc gagacaggaa cgttgctcgt gcccagagta cggagcgatg ccgaggcatt
     2701 gccagatcgg cccgccgggc cccgctgcca ctgcgggacc gcaattgccc acacaccggg
     2761 caaacggccg cgtatctact gctcagaccg ctgccggatg gcagcgaagc gggcgatcgc
     2821 gcgtgtgacg cgagatgccg cccgaggcaa aagcgaacac cttgggaaag aaacaacaga
     2881 gtttcccgca cccctccgac ctgcggtttc tccggacggg gtggatgggg agagcccgag
     2941 aggcgacagc ctctcggaag taggaagcac gtcgcggagc gacgctgccc gactgcggaa
     3001 agccgcccgg tacagccgcc gccggacgct gtggcggatc agcggggacg ccgcgtgcaa
     3061 gggctgcggc cgcgccctga tggaccctgc ctccggcgta atcgtcgccc agacggcggc
     3121 cggaacgtcc gtggtcctgg gcctgatgcg gtgcgggcgg atctggctct gcccggtctg
     3181 cgccgccacg atccggcaca agcgggccga ggagatcacc gccgccgtgg tcgagtggat
     3241 caagcgcggg gggaccgcct acctggtcac cttcacggcc cgccatgggc acacggaccg
     3301 gctcgcggac ctcatggacg ccctccaggg cacccggaag acgccggaca gcccccggcg
     3361 gccgggcgcc taccagcgac tgatcacggg cggcacgtgg gccggacgcc gggccaagga
     3421 cgggcaccgg gccgccgacc gcgagggcat ccgagatcgg atcgggtacg tcggcatgat
     3481 ccgcgcgacc gaagtcaccg tggggcagat caacggctgg cacccgcaca tccacgcgat
     3541 cgtcctggtc ggcggccgga ccgaggggga gcggtccgcg aagcagatcg tcgccacctt
     3601 cgagccgacc ggcgccgcgc tcgacgagtg gcaggggcac tggcggtccg tgtggaccgc
     3661 cgccctgcgc aaggtcaacc ccgccttcac gcccgacgac cggcacggcg tcgacttcaa
     3721 gcggctggaa accgagcgcg acgccaacga cctcgccgag tacatcgcca agacccagga
     3781 cgggaaggcg cccgccctcg aactcgcccg cgccgacctc aagacggcga ccggcgggaa
     3841 cgtcgccccg ttcgaactcc tcggacggat cggggacctg accggcggca tgaccgagga
     3901 cgacgccgcc ggggtcggct cgctggagtg gaacctctcg cgctggcacg agtacgagcg
     3961 ggcaacccgg ggacgccggg ccatcgaatg gacccgctac ctgcggcaga tgctcgggct
     4021 cgacggcggc gacaccgagg ccgacgacct cgatctgctc ctggcggccg acgccgacgg
     4081 cggggagctg cgggccgggg tcgccgtgac cgaggacgga tggcacgcgg tcacccgccg
     4141 cgccctcgac ctcgaggcga cccgggccgc cgaaggcaag gacggcaacg aggattcggc
     4201 ggccgtgggc gaacgggtgc gggaggtcct ggcgctggcc gacgcggccg acacagtggt
     4261 ggtgctcacg gcgggggagg tggccgaggc gtacgccgac atgctcgccg ccctcgccca
     4321 gcgccgcgag gaagcaactg cacgccgacg gcgagagcag gacgacgacc aggacgacga
     4381 cgccgacgac cgccaggagc gggccgcccg gcacatcgcc cggctcgcaa gtgggcccac
     4441 ttcgcactaa ctcgctcccc cccgccgtac gtcatcccgg tgacgtacgg cgggggtcgg
     4501 tgacgtacgc ggcgacggcg gccggggtcg aagccgcggg agtaatcctg ggattactcg
     4561 cccggggtcg gccccgccgg cacttcgtgc aggcggtacc cgct
//
