LOCUS       sequence-398469-        2933 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 2933)
  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 2933)
  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_5
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     296..315
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     380..399
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     509..525
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     509..525
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      526..572
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     686..708
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     700..717
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     701..717
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    943..1446
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(1594..1610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1594..1610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1782..2370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1862..1881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2506..2933,1..388))
                     /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(2506..2933,1..388))
                     /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 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
       61 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
      121 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
      181 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
      241 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
      301 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctcgagc
      361 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
      421 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
      481 gagacacaac gtggctttcc cccgccgctc tagaactagt ggatccaaat aaaacgaaag
      541 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcgcatta tacgagacgt
      601 ccaggttggg atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt
      661 gatccaccac aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat
      721 gcataatccg cacgcatctg gaataaggaa gtgccattcc gcctgacctt agcgtgcgaa
      781 cagcacgcta ttgttgtatc attactcgca tccattctca ggctgtctcg tctcgtctcc
      841 gagctcttca atgagtcagg gtcacgcgta ggacgcgtcg agtagggttc cgacggggag
      901 tatatactgt tccgcagcca cacgccgtgt gaagctgggg agatcacgtg ctataaaaat
      961 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     1021 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     1081 acaaatacta ccttttatct tgctcgtcct gctctcaggt attaatgccg aattgtttca
     1141 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     1201 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     1261 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     1321 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     1381 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     1441 gcgatccgct agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt
     1501 ccacctagcg tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat
     1561 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1621 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1681 tggagttcag ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag
     1741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2221 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2401 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
     2461 aaatcgtccg ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc
     2521 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2581 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2641 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2701 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2761 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2821 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2881 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca acc
//
