LOCUS       sequence-397996-        5344 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 5344)
  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 5344)
  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_3
FEATURES             Location/Qualifiers
     source          1..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     245..264
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     374..390
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     374..390
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      391..437
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     551..573
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     565..582
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     566..582
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3870..3886)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3870..3886)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3883..3905)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(4058..4646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4782..5344,1..253))
                     /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(join(4782..5344,1..253))
                     /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"
ORIGIN
        1 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
       61 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
      121 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
      181 catataaatc agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt
      241 gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc
      301 atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc
      361 tttcccccgc cgctctagaa ctagtggatc caaataaaac gaaaggctca gtcgaaagac
      421 tgggcctttc gttttatctg ttgtttgtcg cattatacga gacgtccagg ttgggatacc
      481 tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata actgtgatcc accacaagcg
      541 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtcatgcata atccgcacgc
      601 atctggaata aggaagtgcc attccgcctg accttagcgt gcgaacagca cgctattgtt
      661 gtatcattac tcgcatccat tctcaggctg tctcgtctcg tctccgagct cttcaatgag
      721 tcagggtcac gcgtaggacg cgtcgagtag ggtttggtca tgtgcttttc gttgttccgc
      781 agccacacgc cgtgtgaagc tggggagccc aagaattaga aatgagtaga tcaaattatt
      841 cacgaataga atcaggaaaa tcagatccaa ccataaaaac actagaacaa attgcaaagt
      901 taactaactc aacgctagta gtggatttaa tcccaaatga gccaacagaa ccagaaccag
      961 aaacagaatc agaacaagta acattggatt tagaaatgga agaagaaaaa agcaatgact
     1021 tcgtgtgaat aatgcacgaa atcgttgctt atttttttta aaagcggtat actagatata
     1081 acgaaacaac gaactgaata gaaacgaaaa aagagccatg acacatttat aaaatgtttg
     1141 acgacatttt ataaatgcat agcccgataa gattgccaaa ccaacgctta tcagttagtc
     1201 agatgaactc ttccctcgta agaagttatt taattaactt tgtttgaaga cggtatataa
     1261 ccgtactatc attatatagg gaaatcagag agttttcaag tatctaagct actgaattta
     1321 agaattgtta agcaatcaat cggaaatcgt ttgattgctt tttttgtatt catttataga
     1381 aggtggagtt tgtatgaatc atgatgaatg taaaacttat ataaaaaata gtttattgga
     1441 gataagaaaa ttagcaaata tctatacact agaaacgttt aagaaagagt tagaaaagag
     1501 aaatatctac ttagaaacaa aatcagataa gtatttttct tcggaggggg aagattatat
     1561 atataagtta atagaaaata acaaaataat ttattcgatt agtggaaaaa aattgactta
     1621 taaaggaaaa aaatcttttt caaaacatgc aatattgaaa cagttgaatg aaaaagcaaa
     1681 ccaagttaat taaacaacct attttatagg atttatagga aaggagaaca gctgaatgaa
     1741 tatccctttt gttgtagaaa ctgtgcttca tgacggcttg ttaaagtaca aatttaaaaa
     1801 tagtaaaatt cgctcaatca ctaccaagcc aggtaaaagc aaaggggcta tttttgcgta
     1861 tcgctcaaaa tcaagcatga ttggcggtcg tggtgttgtt ctgacttccg aggaagcgat
     1921 tcaagaaaat caagatacat ttacacattg gacacccaac gtttatcgtt atggaacgta
     1981 tgcagacgaa aaccgttcat acacgaaagg acattctgaa aacaatttaa gacaaatcaa
     2041 taccttcttt attgattttg atattcacac ggcaaaagaa actatttcag caagcgatat
     2101 tttaacaacc gctattgatt taggttttat gcctactatg attatcaaat ctgataaagg
     2161 ttatcaagca tattttgttt tagaaacgcc agtctatgtg acttcaaaat cagaatttaa
     2221 atctgtcaaa gcagccaaaa taatttcgca aaatatccga gaatattttg gaaagtcttt
     2281 gccagttgat ctaacgtgta atcattttgg tattgctcgc ataccaagaa cggacaatgt
     2341 agaatttttt gatcctaatt accgttattc tttcaaagaa tggcaagatt ggtctttcaa
     2401 acaaacagat aataagggct ttactcgttc aagtctaacg gttttaagcg gtacagaagg
     2461 caaaaaacaa gtagatgaac cctggtttaa tctcttattg cacgaaacga aattttcagg
     2521 agaaaagggt ttaatagggc gtaataacgt catgtttacc ctctctttag cctactttag
     2581 ttcaggctat tcaatcgaaa cgtgcgaata taatatgttt gagtttaata atcgattaga
     2641 tcaaccctta gaagaaaaag aagtaatcaa aattgttaga agtgcctatt cagaaaacta
     2701 tcaaggggct aatagggaat acattaccat tctttgcaaa gcttgggtat caagtgattt
     2761 aaccagtaaa gatttatttg tccgtcaagg gtggtttaaa ttcaagaaaa aaagaagcga
     2821 acgtcaacgt gttcatttgt cagaatggaa agaagattta atggcttata ttagcgaaaa
     2881 aagcgatgta tacaagcctt atttagtgac gaccaaaaaa gagattagag aagtgctagg
     2941 cattcctgaa cggacattag ataaattgct gaaggtactg aaggcgaatc aggaaatttt
     3001 ctttaagatt aaaccaggaa gaaatggtgg cattcaactt gctagtgtta aatcattgtt
     3061 gctatcgatc attaaagtaa aaaaagaaga aaaagaaagc tatataaagg cgctgacaaa
     3121 ttcttttgac ttagagcata cattcattca agagacttta aacaagctag cagaacgccc
     3181 taaaacggac acacaactcg atttgtttag ctatgataca ggctgaaaat aaaacccgca
     3241 ctatgccatt acatttatat ctatgatacg tgtttgtttt ttctttgctg tttagcgaat
     3301 gattagcaga aatatacaga gtaagatttt aattaattat tagggggaga aggagagagt
     3361 agcccgaaaa cttttagttg gcttggactg aacgaagtga gggaaaggct actaaaacgt
     3421 cgaggggcag tgagagcgaa gcgaacactt gattttttaa ttttctatct tttataggtc
     3481 attagagtat acttatttgt cctataaact atttagcagc ataatagatt tattgaatag
     3541 gtcatttaag ttgagcatat tagaggagga aaatcttgga gaaatatttg aagaacccga
     3601 ttacatggat tggattagtt cttgtggtta cgtggttttt aactaaaagt agtgaatttt
     3661 tgatttttgg tgtgtgtgtc ttgttgttag tatttgctag tcaaagtgat taaatagaat
     3721 tccgctagca tgaagagcac cccaggatac atagattacc acaactccga gcccttccac
     3781 ctagcgtgct aaccacgcac gctattgttg tataggctag gtggaggctc agtgatgata
     3841 agtctgcgat ggtggatgca tgtgtcatgg tcatagctgt ttcctgtgtg aaattgttat
     3901 ccgctcagag ggcacaatcc tattccgcgc tatccgacaa tctccaagac attaggtgga
     3961 gttcagttcg gcgtatggca tatgtcgctg gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4201 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4261 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4321 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4381 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4441 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4501 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4561 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4621 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4681 ctctattcaa caaagccgcc gtcccgtcaa gtcagcgtaa atgggtaggg ggcttcaaat
     4741 cgtccgctct gccagtgtta caaccaatta acaaattctg attagaaaaa ctcatcgagc
     4801 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     4861 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     4921 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     4981 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     5041 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     5101 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     5161 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     5221 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     5281 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     5341 tgct
//
