LOCUS       sequence-397958-        4179 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 4179)
  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 4179)
  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_1
FEATURES             Location/Qualifiers
     source          1..4179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(379..1194)
                     /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(379..1194)
                     /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     1102..1121
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1186..1205
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1315..1331
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1315..1331
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      1332..1378
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1492..1514
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1506..1523
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1507..1523
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3646..3662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3659..3681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(3834..4179,1..243))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3914..3933)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
       61 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
      121 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
      181 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
      241 caagaagatc ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc
      301 ccgtcaagtc agcgtaaatg ggtagggggc ttcaaatcgt ccgctctgcc agtgttacaa
      361 ccaattaaca aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
      421 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
      481 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
      541 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
      601 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
      661 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
      721 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
      781 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
      841 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
      901 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
      961 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     1021 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     1081 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     1141 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     1201 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     1261 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcccccg ccgctctaga
     1321 actagtggat ccaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1381 gttgtttgtc gcattatacg agacgtccag gttgggatac ctgaaacaaa acccatcgta
     1441 cggccaagga agtctccaat aactgtgatc caccacaagc gccagggttt tcccagtcac
     1501 gacgttgtaa aacgacggcc agtcatgcat aatccgcacg catctggaat aaggaagtgc
     1561 cattccgcct gaccttagcg tgcgaacagc acgctattgt tgtatcatta ctcgcatcca
     1621 ttctcaggct gtctcgtctc gtctccgagc tcttcaatga gtcagggtca cgcgtaggac
     1681 gcgtcgagta gggtatatag atgccgtcct agcggttccg cagccacacg ccgtgtgaag
     1741 ctggggagag cgtctaccag gacttttacc tgtccgaccc gttgcaacgg aaccccccac
     1801 ggaacccccg cgacacccgc tccccaattg cgttagaaca gcggtggatt gtcggcttcg
     1861 ttgtgggcct tttgagccgc ttcctgttct gccgcacgct ctttcctcgc ccgatagccg
     1921 agtcgcttaa cggtgtccag atgcagcccg aaatgtttgg ccgtttgcgg ccaagagtgg
     1981 ccctcgtcgt cgtgataggc gcggatgcgt tcgcggcgtg cagcctgctc ggcgagccac
     2041 tcgctgcgtt cctgcgccac gagccggacg acgtggcgtt cggatagtcc ggtgattcga
     2101 gcgccttcgg cggcggtcac gcgccgcttt ttgcggacag tcggctgccg gttgtagccg
     2161 tcgctgtagc cgtcgctcat agcaatgcct ccatggctga cgcggacttt gcgcgccgcg
     2221 caactgtgct cgccgccgtg cgcgctgctg cgcccttccg cgagatggcc gactggcgcg
     2281 cactgagtgt ggcctcgtag accacgatcc cgtccgccca aatgcgcgac ttggttgtga
     2341 tccaacgcca aatgctgttg gcgatggcgc ggacctcgct gtccggtagc ggtccgggac
     2401 acacgtcgtt gcacgggaac tcggcgtttc gcgcgtggca ctcggcatag atcgcgcggc
     2461 cgagtccgtc cacgttccgg gtcggcaggt agatccgcat gagggcggga cgataggccc
     2521 acaacctgac ggaatcgaac agtgcgcaat tccgccctag cggcgtcgga gccgctttgt
     2581 acgtggtctg ctgacgccag cgcggcggtg gcatgttcgc gccgagctcg gcctcgatgt
     2641 ggctgagtgt gtagagatct gagtggagcc attccgtttc ccaggcgatg tggccggggt
     2701 ttttggtcat gaggcctgag taactgcggt cgccgtccac ggcgcgccga aggccttcgg
     2761 cgcacgccgc catgtatgcg agcggcttac gccgcgcgta ttcggtgcgt ggaacagggg
     2821 cgttgagtgc ccacactgcg tgtgcgtggc cgttggcgcg attgcccacg atcgcgttgg
     2881 gcagcggatg ggacccccgg gcgctgagcg ctcggagcgc tgcgtctgga tggtctacgt
     2941 ccacgaccag caggtttgcc agcgctgttg ggttcgcctc gatgtaccgg cggcctaggg
     3001 ccgacgcgcg gctttggcgg tagatcccct cgagcagatc gtcgcttgcc agcggccagt
     3061 acggcagcca gagctgctca aattcgtcgg cgacgtggct cacgcttggt agtagaccac
     3121 gattaatcac cggtgtatgg tccgacacga gctccaagtc agatatttcg ctgaggggcc
     3181 accccacaac tgcacactcc cccgctctcc cgtcgagccc tggtggtgga acaccagcga
     3241 cagccgagca cccccaacca cctgtaccaa ccaggaggaa cacatgcgtc gtttcgagga
     3301 cgtttccggg ccgctgagag ccgctgtggc ggccgtacac gccgccttag acccgttaga
     3361 ccccctgccg cctgaatgcg cgggtacgag ccacacagcg cccgaactta cggagctggt
     3421 gggctcacct ggctttatgg cgtacgaatc ggctgtgtgc gacctgttgg gcgaggtgag
     3481 gtacgcgcta ctcacgctcg ctagcatgaa gagcacccca ggatacatag attaccacaa
     3541 ctccgagccc ttccacctag cgtgctaacc acgcacgcta ttgttgtata ggctaggtgg
     3601 aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc cgacaatctc
     3721 caagacatta ggtggagttc agttcggcgt atggcatatg tcgctggaaa gaacatgtga
     3781 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3841 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3901 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3961 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4021 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4081 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4141 cttgagtcca acccggtaag acacgactta tcgccactg
//
