LOCUS       sequence-406191-        4872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4872)
  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 4872)
  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: pGL0_17 [AMA1 short ORI]
FEATURES             Location/Qualifiers
     source          1..4872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      564..1152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1053..1072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1306..1323
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4062..4080)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4148..4252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4474..4872,1..393)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
       61 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
      121 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
      181 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
      241 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
      301 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccattacgta
      361 aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag accaagttta ctcatatata
      421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      541 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      601 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      661 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      721 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      781 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      841 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      901 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      961 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1021 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1081 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1141 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1201 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1261 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     1321 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     1381 taatcactct gtggtctcag gagcagtatg cttgatgtta tccaaagtgt gagagaggaa
     1441 ggtccaaaca tacacgattg ggagagggcc taggtataag agtttttgag tagaacgcat
     1501 gtgagcccag ccatctcgag gagattaaac acgggccggc atttgatggc tatgttagta
     1561 ccccaatgga aagcctgaga gtccagtggt cgcagataac tccctaaatt ccctgagcta
     1621 actctaagtc gaccatgccg tttatggtta gcgcctccca aaaaggaatg gccgacttag
     1681 agttacctct tgaccgactt tttctttcct cccccttaca tttcgttacc acaacacatt
     1741 cctatatcca aactccaggt acataactag tcgaaatctc ttttaaatct agtcaagaac
     1801 tagatactta accttcatac tgcttagaga tagcctttga agctcttatt tcgctctttg
     1861 tctcactctc accttcctcc tcctcctcca atcctttttg ctcgggacta gtccaagact
     1921 agtcaacgat tagtatgccg agcattcgcg ataacgatct ccgaaagtcc ccagagtact
     1981 gtcactatct cgaggcagtt aaggacgggg agcttacgct gccggatttc aagatagtaa
     2041 gccgacccga ctagctttaa gcctagttac tgactagtcc acaggacgac aatggcgtgc
     2101 ctgatatcca tccatatgaa gtctactgcc gagtgaaggg atgcctcaag cgtacagtga
     2161 gtctactgct atactagttt ctgactagtt attaactagt tgacaggttc cctctgccaa
     2221 cagaaacata ttggtcaagc acttgaagga caagaactcc cacggcatgg agtttacatt
     2281 gcacaatgga cctcccacta tgaaggaact gatggaggcc aaaggcaagt cctatctaga
     2341 ttacttggtg actagtccct gactagtcaa gtagcatggt atgaaggctt gtttgaaggc
     2401 actgttctcc caaccccgac tcctaccaag aagcgcaagc gagctgcgta agtttctgtg
     2461 agtctaacta gtgtattagc taatatatag cagcaccaag tccaaggacc acaatactaa
     2521 gggtgtcgag aattcgtgag tttcttctcc catttcaact agtccttgac tagtcactaa
     2581 ctactttgca gcaacgaggg tgaagctgga aatgatcagg acaatggcga gggcccgtaa
     2641 gtacagccat tcaatgcaga ctagttgcta actaatctgt gactagttca agtggtccgt
     2701 acgccgtgca tacccctgtg actggtagga atttgagcaa gcctgtcttg ccgcgcgatg
     2761 agaaaggaaa ggcaagttac attcagcccc gtacctagga tcagtgctaa tttataacct
     2821 ctagccactc tttatgcaga tccgccgtga gggtagcaag gcagctaaat cagctggtga
     2881 gaaaggaacc ataccctgca agacctgtcg caacgcaaag ggcaaaggta agctatccaa
     2941 gctagtttgg gactagattc taactagtct cagcaccgtg tggttcaaag ccatattgcg
     3001 agttttggcg ctttttctca tcgattgacg aggcaaaggg agcgagtatg caacctcgta
     3061 agtcagactc agacaaagac caactagtta ttgaccagtc atctagaagg ctctgttgtg
     3121 gatcttgagg ccctggagag ttcctccaac aatccggaga caagcaagtc ctcgtcggac
     3181 tagtcactaa ctagactcta actagttgca gacatggata atgcaaaaga gacaagcaat
     3241 gaagaaagtg gtaagacatt tctcctttgt ggttctggac tagtctttga ctagtcacag
     3301 tcttaaacaa ggaaaatgag catgaaaatg aggaggaaaa ggctgctgag cccgaggaag
     3361 tgcagggtga tggcagacat ggtaggttaa taccttgtta gttattgcta gtcactgact
     3421 agtcaataac tagtctctga acaccttgca atcactccgt ttgcgcagct gaacagtggt
     3481 gaggataata gtagtaagtt attctagctt cagagttata ggagactaga tactaactag
     3541 tattagttgc aactaacctg gatctcagag actttggcct caatctagaa tctatctagt
     3601 tgtcaactag actgtggtat cattgtcttt tattttccta gtcctggaac tagcttctaa
     3661 ctagtctccc taatatgtgg ctgtcttgtt tttttttttt gtttccctac ccggatatct
     3721 agtccccttc taggttctgt taacctctcg ggctctgatt tagtttaacg caaacctgag
     3781 attagtttct aactagtctc taggttttct atccaccttt aattgtaata ataaatacaa
     3841 gcaacgttta tacgtcaaaa gcatttataa acttttaccc taaagtagct tgcttgtgtg
     3901 tttagtttat aattagtctc ttattaattt gatgtaggta agcccgccac aaatatatat
     3961 ttttaacaag ataccgtgga aaaacttcgt gctatcacaa aacagtatac aaaaaataag
     4021 ctcgcttgag accacgaagt ggctcttcag tggacgaaag ggcctcgtga tacgcctatt
     4081 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4141 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4201 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgcgctc
     4261 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     4321 catggcttgt tatgactgtt tttttttggg gtacagtcta tgcctcgggc atccaagcag
     4381 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     4441 cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat cgccgaagta
     4501 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     4561 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag cgatattgat
     4621 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     4681 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     4741 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     4801 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     4861 gatctggcta tc
//
