LOCUS       sequence-398470-        5048 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 5048)
  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 5048)
  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_6
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..170
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     235..254
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     364..380
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     364..380
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      381..427
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     541..563
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     555..572
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     556..572
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3564..3580)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3564..3580)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3577..3599)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3752..4340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3832..3851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4476..5048,1..243))
                     /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(4476..5048,1..243))
                     /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 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
       61 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
      121 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
      181 agcatccatg ttggaattta atcgcggcct cgagcaagac gtttcccgtt gaatatggct
      241 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
      301 atttttatct tgtgcaatgt aacatcagag attttgagac acaacgtggc tttcccccgc
      361 cgctctagaa ctagtggatc caaataaaac gaaaggctca gtcgaaagac tgggcctttc
      421 gttttatctg ttgtttgtcg cattatacga gacgtccagg ttgggatacc tgaaacaaaa
      481 cccatcgtac ggccaaggaa gtctccaata actgtgatcc accacaagcg ccagggtttt
      541 cccagtcacg acgttgtaaa acgacggcca gtcatgcata atccgcacgc atctggaata
      601 aggaagtgcc attccgcctg accttagcgt gcgaacagca cgctattgtt gtatcattac
      661 tcgcatccat tctcaggctg tctcgtctcg tctccgagct cttcaatgag tcagggtcac
      721 gcgtaggacg cgtcgagtag ggtcatgttt aggaacgcta ccggttccgc agccacacgc
      781 cgtgtgaagc tggggagcag tatgcttgat gttatccaaa gtgtgagaga ggaaggtcca
      841 aacatacacg attgggagag ggcctaggta taagagtttt tgagtagaac gcatgtgagc
      901 ccagccatct cgaggagatt aaacacgggc cggcatttga tggctatgtt agtaccccaa
      961 tggaaagcct gagagtccag tggtcgcaga taactcccta aattccctga gctaactcta
     1021 agtcgaccat gccgtttatg gttagcgcct cccaaaaagg aatggccgac ttagagttac
     1081 ctcttgaccg actttttctt tcctccccct tacatttcgt taccacaaca cattcctata
     1141 tccaaactcc aggtacataa ctagtcgaaa tctcttttaa atctagtcaa gaactagata
     1201 cttaaccttc atactgctta gagatagcct ttgaagctct tatttcgctc tttgtctcac
     1261 tctcaccttc ctcctcctcc tccaatcctt tttgctcggg actagtccaa gactagtcaa
     1321 cgattagtat gccgagcatt cgcgataacg atctccgaaa gtccccagag tactgtcact
     1381 atctcgaggc agttaaggac ggggagctta cgctgccgga tttcaagata gtaagccgac
     1441 ccgactagct ttaagcctag ttactgacta gtccacagga cgacaatggc gtgcctgata
     1501 tccatccata tgaagtctac tgccgagtga agggatgcct caagcgtaca gtgagtctac
     1561 tgctatacta gtttctgact agttattaac tagttgacag gttccctctg ccaacagaaa
     1621 catattggtc aagcacttga aggacaagaa ctcccacggc atggagttta cattgcacaa
     1681 tggacctccc actatgaagg aactgatgga ggccaaaggc aagtcctatc tagattactt
     1741 ggtgactagt ccctgactag tcaagtagca tggtatgaag gcttgtttga aggcactgtt
     1801 ctcccaaccc cgactcctac caagaagcgc aagcgagctg cgtaagtttc tgtgagtcta
     1861 actagtgtat tagctaatat atagcagcac caagtccaag gaccacaata ctaagggtgt
     1921 cgagaattcg tgagtttctt ctcccatttc aactagtcct tgactagtca ctaactactt
     1981 tgcagcaacg agggtgaagc tggaaatgat caggacaatg gcgagggccc gtaagtacag
     2041 ccattcaatg cagactagtt gctaactaat ctgtgactag ttcaagtggt ccgtacgccg
     2101 tgcatacccc tgtgactggt aggaatttga gcaagcctgt cttgccgcgc gatgagaaag
     2161 gaaaggcaag ttacattcag ccccgtacct aggatcagtg ctaatttata acctctagcc
     2221 actctttatg cagatccgcc gtgagggtag caaggcagct aaatcagctg gtgagaaagg
     2281 aaccataccc tgcaagacct gtcgcaacgc aaagggcaaa ggtaagctat ccaagctagt
     2341 ttgggactag attctaacta gtctcagcac cgtgtggttc aaagccatat tgcgagtttt
     2401 ggcgcttttt ctcatcgatt gacgaggcaa agggagcgag tatgcaacct cgtaagtcag
     2461 actcagacaa agaccaacta gttattgacc agtcatctag aaggctctgt tgtggatctt
     2521 gaggccctgg agagttcctc caacaatccg gagacaagca agtcctcgtc ggactagtca
     2581 ctaactagac tctaactagt tgcagacatg gataatgcaa aagagacaag caatgaagaa
     2641 agtggtaaga catttctcct ttgtggttct ggactagtct ttgactagtc acagtcttaa
     2701 acaaggaaaa tgagcatgaa aatgaggagg aaaaggctgc tgagcccgag gaagtgcagg
     2761 gtgatggcag acatggtagg ttaatacctt gttagttatt gctagtcact gactagtcaa
     2821 taactagtct ctgaacacct tgcaatcact ccgtttgcgc agctgaacag tggtgaggat
     2881 aatagtagta agttattcta gcttcagagt tataggagac tagatactaa ctagtattag
     2941 ttgcaactaa cctggatctc agagactttg gcctcaatct agaatctatc tagttgtcaa
     3001 ctagactgtg gtatcattgt cttttatttt cctagtcctg gaactagctt ctaactagtc
     3061 tccctaatat gtggctgtct tgtttttttt ttttgtttcc ctacccggat atctagtccc
     3121 cttctaggtt ctgttaacct ctcgggctct gatttagttt aacgcaaacc tgagattagt
     3181 ttctaactag tctctaggtt ttctatccac ctttaattgt aataataaat acaagcaacg
     3241 tttatacgtc aaaagcattt ataaactttt accctaaagt agcttgcttg tgtgtttagt
     3301 ttataattag tctcttatta atttgatgta ggtaagcccg ccacaaatat atatttttaa
     3361 caagataccg tggaaaaact tcgtgctatc acaaaacagt atacaaaaaa taagctcgct
     3421 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
     3481 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
     3541 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3601 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
     3661 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
     3721 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3781 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3841 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3901 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3961 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4021 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4081 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4141 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4201 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4261 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
     4381 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     4441 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     4501 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     4561 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     4621 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     4681 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     4741 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     4801 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     4861 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     4921 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     4981 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     5041 atggtcgg
//
