LOCUS       sequence-397972-        4575 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 4575)
  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 4575)
  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_4
FEATURES             Location/Qualifiers
     source          1..4575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..95
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     205..221
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      222..268
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     382..404
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     396..413
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     397..413
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2932..2948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2932..2948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2945..2967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3120..3708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3200..3219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3844..4575,1..84))
                     /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(3844..4575,1..84))
                     /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     join(4567..4575,1..11)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgagcaaga
       61 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
      121 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
      181 cacaacgtgg ctttcccccg ccgctctaga actagtggat ccaaataaaa cgaaaggctc
      241 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc gcattatacg agacgtccag
      301 gttgggatac ctgaaacaaa acccatcgta cggccaagga agtctccaat aactgtgatc
      361 caccacaagc gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtcatgcat
      421 aatccgcacg catctggaat aaggaagtgc cattccgcct gaccttagcg tgcgaacagc
      481 acgctattgt tgtatcatta ctcgcatcca ttctcaggct gtctcgtctc gtctccgagc
      541 tcttcaatga gtcagggtca cgcgtaggac gcgtcgagta gggtgggtgg gtaaatggta
      601 atgcgttccg cagccacacg ccgtgtgaag ctggggagaa ctaactcaac gctagtagtg
      661 gatttaatcc caaatgagcc aacagaacca gaaccagaaa cagaacaagt aacattggag
      721 ttagaaatgg aagaagaaaa aagcaatgat ttcgtgtgaa taatgcacga aatcattgct
      781 tattttttta aaaagcgata tactagatat aacgaaacaa cgaactgaat aaagaataca
      841 aaaaaagagc cacgaccagt taaagcctga gaaattttaa ctgcgagcct taattgatta
      901 ccaccaatca attaaagaag tccagaccca aaatttggta aagtatttaa ttactttatt
      961 aatcagatac ttaaatatct gtaaacccat tatatcgggt ttttgagggg atttcaagtc
     1021 tttaagaaga taccaggcaa tcaattaaga aaaacttagt tgattgcctt ttttgttgtg
     1081 attcaacttt gatcgtagct tctaactaat taattttcgt aagaaaggag aacagctgaa
     1141 tgaatatccc ttttgttgta gaaactgtgc ttcatgacgg cttgttaaag tacaaattta
     1201 aaaatagtaa aattcgctca atcactacca agccaggtaa aagtaaaggg gctatttttg
     1261 cgtatcgctc aaaaaaaagc atgattggcg gacgtggcgt tgttctgact tccgaagaag
     1321 cgattcacga aaatcaagat acatttacgc attggacacc aaacgtttat cgttatggta
     1381 cgtatgcaga cgaaaaccgt tcatacacta aaggacattc tgaaaacaat ttaagacaaa
     1441 tcaatacctt ctttattgat tttgatattc acacggaaaa agaaactatt tcagcaagcg
     1501 atattttaac aacagctatt gatttaggtt ttatgcctac gttaattatc aaatctgata
     1561 aaggttatca agcatatttt gttttagaaa cgccagtcta tgtgacttca aaatcagaat
     1621 ttaaatctgt caaagcagcc aaaataatct cgcaaaatat ccgagaatat tttggaaagt
     1681 ctttgccagt tgatctaacg tgcaatcatt ttgggattgc tcgtatacca agaacggaca
     1741 atgtagaatt ttttgatccc aattaccgtt attctttcaa agaatggcaa gattggtctt
     1801 tcaaacaaac agataataag ggctttactc gttcaagtct aacggtttta agcggtacag
     1861 aaggcaaaaa acaagtagat gaaccctggt ttaatctctt attgcacgaa acgaaatttt
     1921 caggagaaaa gggtttagta gggcgcaata gcgttatgtt taccctctct ttagcctact
     1981 ttagttcagg ctattcaatc gaaacgtgcg aatataatat gtttgagttt aataatcgat
     2041 tagatcaacc cttagaagaa aaagaagtaa tcaaaattgt tagaagtgcc tattcagaaa
     2101 actatcaagg ggctaatagg gaatacatta ccattctttg caaagcttgg gtatcaagtg
     2161 atttaaccag taaagattta tttgtccgtc aagggtggtt taaattcaag aaaaaaagaa
     2221 gcgaacgtca acgtgttcat ttgtcagaat ggaaagaaga tttaatggct tatattagcg
     2281 aaaaaagcga tgtatacaag ccttatttag cgacgaccaa aaaagagatt agagaagtgc
     2341 taggcattcc tgaacggaca ttagataaat tgctgaaggt actgaaggcg aatcaggaaa
     2401 ttttctttaa gattaaacca ggaagaaatg gtggcattca acttgctagt gttaaatcat
     2461 tgttgctatc gatcattaaa ttaaaaaaag aagaacgaga aagctatata aaggcgctga
     2521 cagcttcgtt taatttagaa cgtacattta ttcaagaaac tctaaacaaa ttggcagaac
     2581 gccccaaaac ggacccacaa ctcgatttgt ttagctacga tacaggctga aaataaaacc
     2641 cgcactatgc cattacattt atatctatga tacgtgtttg tttttctttg ctgtttagtg
     2701 aatgattagc agaaatatac agagtaagat tttaattaat tattaggggg agaaggagag
     2761 agtagcccga aaacttttag ttggcgctag catgaagagc accccaggat acatagatta
     2821 ccacaactcc gagcccttcc acctagcgtg ctaaccacgc acgctattgt tgtataggct
     2881 aggtggaggc tcagtgatga taagtctgcg atggtggatg catgtgtcat ggtcatagct
     2941 gtttcctgtg tgaaattgtt atccgctcag agggcacaat cctattccgc gctatccgac
     3001 aatctccaag acattaggtg gagttcagtt cggcgtatgg catatgtcgc tggaaagaac
     3061 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3121 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3181 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3241 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3301 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3361 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3421 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3481 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3541 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3601 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3661 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3721 atcttttcta cggggtctga cgctctattc aacaaagccg ccgtcccgtc aagtcagcgt
     3781 aaatgggtag ggggcttcaa atcgtccgct ctgccagtgt tacaaccaat taacaaattc
     3841 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     3901 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3961 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     4021 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     4081 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     4141 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     4201 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     4261 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     4321 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     4381 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     4441 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     4501 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     4561 gacattatcg cgagc
//
