LOCUS       sequence-400885-        7729 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 7729)
  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 7729)
  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_59
FEATURES             Location/Qualifiers
     source          1..7729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(144..160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(157..179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(332..920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(412..431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1056..1871)
                     /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(1056..1871)
                     /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     1779..1798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1863..1882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1992..2008
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1992..2008
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2009..2055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2169..2191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2183..2200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2184..2200
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2434..7682
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
ORIGIN
        1 agcatgaaga gcaccccagg atacatagat taccacaact ccgagccctt ccacctagcg
       61 tgctaaccac gcacgctatt gttgtatagg ctaggtggag gctcagtgat gataagtctg
      121 cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
      181 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
      241 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
      301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
      361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
      421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
      481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
      541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
      601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
      661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
      721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
      781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
      841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
      901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
      961 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtccg
     1021 ctctgccagt gttacaacca attaacaaat tctgattaga aaaactcatc gagcatcaaa
     1081 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1141 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1201 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1261 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     1321 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1381 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1441 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1501 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     1561 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     1621 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     1681 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     1741 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     1801 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     1861 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     1921 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     1981 tcccccgccg ctctagaact agtggatcca aataaaacga aaggctcagt cgaaagactg
     2041 ggcctttcgt tttatctgtt gtttgtcgca ttatacgaga cgtccaggtt gggatacctg
     2101 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     2161 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     2221 ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg ctattgttgt
     2281 atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgagctct tcaatgagtc
     2341 agggtcacgc gtaggacgcg tcgagtaggg ttaaatatcc ccttgaggcc cgttccgcag
     2401 ccacacgccg tgtgaagctg gggagagctt gatagcttat tttttgtata ctgttttgtg
     2461 atagcacgaa gtttttccac ggtatcttgt taaaaatata tatttgtggc gggcttacct
     2521 acatcaaatt aataagagac taattataaa ctaaacacac aagcaagcta ctttagggta
     2581 aaagtttata aatgcttttg acgtataaac gttgcttgta tttattatta caattaaagg
     2641 tggatagaaa acctagagac tagttagaaa ctaatctcag gtttgcgtta aactaaatca
     2701 gagcccgaga ggttaacaga acctagaagg ggactagata tccgggtagg gaaacaaaaa
     2761 aaaaaaacaa gacagccaca tattagggag actagttaga agctagttcc aggactagga
     2821 aaataaaaga caatgatacc acagtctagt tgacaactag atagattcta gattgaggcc
     2881 aaagtctctg agatccaggt tagttgcaac taatactagt tagtatctag tctcctataa
     2941 ctctgaagct agaataactt actactatta tcctcaccac tgttcagctg cgcaaacgga
     3001 gtgattgcaa ggtgttcaga gactagttat tgactagtca gtgactagca ataactaaca
     3061 aggtattaac ctaccatgtc tgccatcacc ctgcacttcc tcgggctcag cagccttttc
     3121 ctcctcattt tcatgctcat tttccttgtt taagactgtg actagtcaaa gactagtcca
     3181 gaaccacaaa ggagaaatgt cttaccactt tcttcattgc ttgtctcttt tgcattatcc
     3241 atgtctgcaa ctagttagag tctagttagt gactagtccg acgaggactt gcttgtctcc
     3301 ggattgttgg aggaactctc cagggcctca agatccacaa cagagccttc tagaagactg
     3361 gtcaataact agttggtctt tgtctgagtc tgacttacga ggttgcatac tcgctccctt
     3421 tgcctcgtca atcgatgaga aaaagcgcca aaactcgcaa tatggctttg aaccacacgg
     3481 tgctgagact agttagaatc tagtcccaaa ctagcttgga tagcttacct ttgccctttg
     3541 cgttgcgaca ggtcttgcag ggtatggttc ctttctcacc agctgattta gctgccttgc
     3601 taccctcacg gcggatctgc ataaagagtg gctagaggtt ataaattagc actgatccta
     3661 ggtacggggc tgaatgtaac ttgcctttcc tttctcatcg cgcggcaaga caggcttgct
     3721 caaattccta ccagtcacag gggtatgcac ggcgtacgga ccacttgaac tagtcacaga
     3781 ttagttagca actagtctgc attgaatggc tgtacttacg ggccctcgcc attgtcctga
     3841 tcatttccag cttcaccctc gttgctgcaa agtagttagt gactagtcaa ggactagttg
     3901 aaatgggaga agaaactcac gaattctcga cacccttagt attgtggtcc ttggacttgg
     3961 tgctgctata tattagctaa tacactagtt agactcacag aaacttacgc agctcgcttg
     4021 cgcttcttgg taggagtcgg ggttgggaga acagtgcctt caaacaagcc ttcataccat
     4081 gctacttgac tagtcaggga ctagtcacca agtaatctag ataggacttg cctttggcct
     4141 ccatcagttc cttcatagtg ggaggtccat tgtgcaatgt aaactccatg ccgtgggagt
     4201 tcttgtcctt caagtgcttg accaatatgt ttctgttggc agagggaacc tgtcaactag
     4261 ttaataacta gtcagaaact agtatagcag tagactcact gtacgcttga ggcatccctt
     4321 cactcggcag tagacttcat atggatggat atcaggcacg ccattgtcgt cctgtggact
     4381 agtcagtaac taggcttaaa gctagtcggg tcggcttact atcttgaaat ccggcagcgt
     4441 aagctccccg tccttaactg cctcgagata gtgacagtac tctggggact ttcggagatc
     4501 gttatcgcga atgctcggca tactaatcgt tgactagtct tggactagtc ccgagcaaaa
     4561 aggattggag gaggaggagg aaggtgagag tgagacaaag agcgaaataa gagcttcaaa
     4621 ggctatctct aagcagtatg aaggttaagt atctagttct tgactagatt taaaagagat
     4681 ttcgactagt tatgtacctg gagtttggat ataggaatgt gttgtggtaa cgaaatgtaa
     4741 gggggaggaa agaaaaagtc ggtcaagagg taactctaag tcggccattc ctttttggga
     4801 ggcgctaacc ataaacggca tggtcgactt agagttagct cagggaattt agggagttat
     4861 ctgcgaccac cgaggaacgg cggaatgcca aagaatcccg atggagctct agctggcggt
     4921 tgacaacccc accttttggc gtttctgcgg cgttgcaggc gggactggat acttcgtaga
     4981 accagaaagg caaggcagaa cgcgctcagc aagagtgttg gaagtgatag catgatgtgc
     5041 cttgttaact aggtcaaaat ctgcagtatg cttgatgtta tccaaagtgt gagagaggaa
     5101 ggtccaaaca tacacgattg ggagagggcc taggtataag agtttttgag tagaacgcat
     5161 gtgagcccag ccatctcgag gagattaaac acgggccggc atttgatggc tatgttagta
     5221 ccccaatgga aagcctgaga gtccagtggt cgcagataac tccctaaatt ccctgagcta
     5281 actctaagtc gaccatgccg tttatggtta gcgcctccca aaaaggaatg gccgacttag
     5341 agttacctct tgaccgactt tttctttcct cccccttaca tttcgttacc acaacacatt
     5401 cctatatcca aactccaggt acataactag tcgaaatctc ttttaaatct agtcaagaac
     5461 tagatactta accttcatac tgcttagaga tagcctttga agctcttatt tcgctctttg
     5521 tctcactctc accttcctcc tcctcctcca atcctttttg ctcgggacta gtccaagact
     5581 agtcaacgat tagtatgccg agcattcgcg ataacgatct ccgaaagtcc ccagagtact
     5641 gtcactatct cgaggcagtt aaggacgggg agcttacgct gccggatttc aagatagtaa
     5701 gccgacccga ctagctttaa gcctagttac tgactagtcc acaggacgac aatggcgtgc
     5761 ctgatatcca tccatatgaa gtctactgcc gagtgaaggg atgcctcaag cgtacagtga
     5821 gtctactgct atactagttt ctgactagtt attaactagt tgacaggttc cctctgccaa
     5881 cagaaacata ttggtcaagc acttgaagga caagaactcc cacggcatgg agtttacatt
     5941 gcacaatgga cctcccacta tgaaggaact gatggaggcc aaaggcaagt cctatctaga
     6001 ttacttggtg actagtccct gactagtcaa gtagcatggt atgaaggctt gtttgaaggc
     6061 actgttctcc caaccccgac tcctaccaag aagcgcaagc gagctgcgta agtttctgtg
     6121 agtctaacta gtgtattagc taatatatag cagcaccaag tccaaggacc acaatactaa
     6181 gggtgtcgag aattcgtgag tttcttctcc catttcaact agtccttgac tagtcactaa
     6241 ctactttgca gcaacgaggg tgaagctgga aatgatcagg acaatggcga gggcccgtaa
     6301 gtacagccat tcaatgcaga ctagttgcta actaatctgt gactagttca agtggtccgt
     6361 acgccgtgca tacccctgtg actggtagga atttgagcaa gcctgtcttg ccgcgcgatg
     6421 agaaaggaaa ggcaagttac attcagcccc gtacctagga tcagtgctaa tttataacct
     6481 ctagccactc tttatgcaga tccgccgtga gggtagcaag gcagctaaat cagctggtga
     6541 gaaaggaacc ataccctgca agacctgtcg caacgcaaag ggcaaaggta agctatccaa
     6601 gctagtttgg gactagattc taactagtct cagcaccgtg tggttcaaag ccatattgcg
     6661 agttttggcg ctttttctca tcgattgacg aggcaaaggg agcgagtatg caacctcgta
     6721 agtcagactc agacaaagac caactagtta ttgaccagtc ttctagaagg ctctgttgtg
     6781 gatcttgagg ccctggagag ttcctccaac aatccggaga caagcaagtc ctcgtcggac
     6841 tagtcactaa ctagactcta actagttgca gacatggata atgcaaaaga gacaagcaat
     6901 gaagaaagtg gtaagacatt tctcctttgt ggttctggac tagtctttga ctagtcacag
     6961 tcttaaacaa ggaaaatgag catgaaaatg aggaggaaaa ggctgctgag cccgaggaag
     7021 tgcagggtga tggcagacat ggtaggttaa taccttgtta gttattgcta gtcactgact
     7081 agtcaataac tagtctctga acaccttgca atcactccgt ttgcgcagct gaacagtggt
     7141 gaggataata gtagtaagtt attctagctt cagagttata ggagactaga tactaactag
     7201 tattagttgc aactaacctg gatctcagag actttggcct caatctagaa tctatctagt
     7261 tgtcaactag actgtggtat cattgtcttt tattttccta gtcctggaac tagcttctaa
     7321 ctagtctccc taatatgtgg ctgtcttgtt tttttttttt gtttccctac ccggatatct
     7381 agtccccttc taggttctgt taacctctcg ggctctgatt tagtttaacg caaacctgag
     7441 attagtttct aactagtctc taggttttct atccaccttt aattgtaata ataaatacaa
     7501 gcaacgttta tacgtcaaaa gcatttataa acttttaccc taaagtagct tgcttgtgtg
     7561 tttagtttat aattagtctc ttattaattt gatgtaggta agcccgccac aaatatatat
     7621 ttttaacaag ataccgtgga aaaacttcgt gctatcacaa aacagtatac aaaaaataag
     7681 ctatcgaatt cctgcagccc cagatcatcc tgtcttcagt cttaacgct
//
