LOCUS       sequence-398477-        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_8
FEATURES             Location/Qualifiers
     source          1..2881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     458..477
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     542..561
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     671..687
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     671..687
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      688..734
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     848..870
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     862..879
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     863..879
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1717..1739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1892..2480)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1972..1991)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2616..2881,1..550))
                     /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(2616..2881,1..550))
                     /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 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
       61 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
      121 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
      181 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
      241 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
      301 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
      361 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
      421 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
      481 ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa
      541 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
      601 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
      661 cccccgccgc tctagaacta gtggatccaa ataaaacgaa aggctcagtc gaaagactgg
      721 gcctttcgtt ttatctgttg tttgtcgcat tatacgagac gtccaggttg ggatacctga
      781 aacaaaaccc atcgtacggc caaggaagtc tccaataact gtgatccacc acaagcgcca
      841 gggttttccc agtcacgacg ttgtaaaacg acggccagtc atgcataatc cgcacgcatc
      901 tggaataagg aagtgccatt ccgcctgacc ttagcgtgcg aacagcacgc tattgttgta
      961 tcattactcg catccattct caggctgtct cgtctcgtct ccgagctctt caatgagtca
     1021 gggtcacgcg taggacgcgt cgagtagggt ggcgagagga gatatagagc gttccgcagc
     1081 cacacgccgt gtgaagctgg ggagtcaaca tctttggata atatcagaat gagaaagaac
     1141 agatacgcag tacgtttttt ggtgagctct ttgcacttct ttagttcttt ccatcaatat
     1201 cagtttttta aacttttagg actaaaagtg atgtttaact tcaaaatgtt taaaattttg
     1261 ttcttcccga cgttcattaa gaatactaat acactttaat aattatttta atattttgtt
     1321 ctaaataatg acttttaatt aaaaaagata aaatataaaa acatcataat aactcaccag
     1381 aggttaagaa caaaaaaaca aattagatat ctgctaatcc aatatagtta aatcaatctt
     1441 tccttggtat aatgggtata ttacatatat ttcaaggacc gacactccta ccaaatatct
     1501 aaaatttacc atattaacat aacatgtata taaacgtcaa atcataatca gcactacgct
     1561 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
     1621 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
     1681 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1741 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
     1801 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
     1861 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1921 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1981 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2041 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2101 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2161 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2221 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2281 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2341 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2401 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2461 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
     2521 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     2581 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     2641 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2701 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2761 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2821 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     2881 t
//
