LOCUS       sequence-398473-        2881 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 2881)
  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 2881)
  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_7
FEATURES             Location/Qualifiers
     source          1..2881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     422..441
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     506..525
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     635..651
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     635..651
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      652..698
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     812..834
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     826..843
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     827..843
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1668..1684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1668..1684)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1681..1703)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1856..2444)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1936..1955)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2580..2881,1..514))
                     /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(2580..2881,1..514))
                     /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 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg
       61 cctgagcgag acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat
      121 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt
      181 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
      241 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
      301 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
      361 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat
      421 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
      481 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
      541 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
      601 gattttgaga cacaacgtgg ctttcccccg ccgctctaga actagtggat ccaaataaaa
      661 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc gcattatacg
      721 agacgtccag gttgggatac ctgaaacaaa acccatcgta cggccaagga agtctccaat
      781 aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      841 agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct gaccttagcg
      901 tgcgaacagc acgctattgt tgtatcatta ctcgcatcca ttctcaggct gtctcgtctc
      961 gtctccgagc tcttcaatga gtcagggtca cgcgtaggac gcgtcgagta gggttaacta
     1021 ccagtatcca ccgcgttccg cagccacacg ccgtgtgaag ctggggagtc aacatctttg
     1081 gataatatca gaatgagaaa gaacagatac gcagtacgtt ttttggtgag ctctttgcac
     1141 ttctttagtt ctttccatca atatcagttg cttatgcact tatgactaat attgatgttt
     1201 aacttcaata tctttaaact tttgttcttc ccgacgttca ttaagaatac taatacactt
     1261 taataattag tttaatattt gtttctatat aatgacattt aattaaaaaa gataaaatat
     1321 aaaaacatca taataactca ccagaggtta agaacaaaaa aacaaattag atatctgcta
     1381 atccaatata gttaaatcaa tctttccttg gtataatggg tatattacat atatttcaag
     1441 gaccgacact cctaccaaat atctaaaatt taccatatta acataacatg tatataaacg
     1501 tcaaatcata atcagcacta cgctagcatg aagagcaccc caggatacat agattaccac
     1561 aactccgagc ccttccacct agcgtgctaa ccacgcacgc tattgttgta taggctaggt
     1621 ggaggctcag tgatgataag tctgcgatgg tggatgcatg tgtcatggtc atagctgttt
     1681 cctgtgtgaa attgttatcc gctcagaggg cacaatccta ttccgcgcta tccgacaatc
     1741 tccaagacat taggtggagt tcagttcggc gtatggcata tgtcgctgga aagaacatgt
     1801 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1861 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1921 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1981 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2041 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2101 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2161 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2221 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2281 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2341 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2401 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2461 tttctacggg gtctgacgct ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat
     2521 gggtaggggg cttcaaatcg tccgctctgc cagtgttaca accaattaac aaattctgat
     2581 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     2641 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2701 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2761 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2821 gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag
     2881 c
//
