LOCUS       sequence-417256-        5881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains two plasmids. Constitutive expression of BsmAIM and 
            inducible expression of BsaIR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  AUTHORS   Bhadra S, Nguyen V, Torres JA, Kar S, Fadanka S, Gandini C, Akligoh 
            H, Paik I, Maranhao AC, Molloy J, Ellington AD
  TITLE     Producing molecular biology reagents without purification.
  JOURNAL   PLoS One. 2021 Jun 1;16(6):e0252507. doi: 
            10.1371/journal.pone.0252507. eCollection 2021.
  PUBMED    34061896
REFERENCE   2  (bases 1 to 5881)
  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: "PLoS One.";
            date: "2021-06-1"; volume: "16(6):e0252507. doi"; pages: " 
            10.1371/journal.pone.0252507. eCollection 2021"
COMMENT     Sequence Label: pBR BsmAIM Kan + p15A BsaIR Amp
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(3307..3326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3517..3538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3992..4786
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4046..4065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4656..4675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      5112..5700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5601..5620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5854..5871
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga acggaggagc ttgcgtcaag
      241 gaaaatacgg agattaatat cgacattaag aaagctgcgc tttgggatac tattcgtaac
      301 aaaagtcagt tcctggagac ccaaatggat ccgcttgagc gcaagcgcac gggctcatac
      361 tttacggcgt tagaactgac tgacgtgatg atgcaggaat tggtcagtta catccttaaa
      421 agtgataaag atatcacgga attgaaattt ctggagccgt gtgtagggac ggggaacttc
      481 gtattttcat atcttaagga aattagtaaa ttgcaactgc acaaagaaca gatcgaaaca
      541 ctgatcaata atatttacgt tgcagacatc aaccaaacgg ctttgttaga gtacaaaaaa
      601 cttcttagca aattcgcaaa actgtacttc gacattgatt tgtccgaaga gtactttaac
      661 tcacacattg gatccgctct gttaattgac gtcgccgcag aacaaccgga gtacattaag
      721 attacggatg tatttcccga tgaggtagtt aaggaaggtt ttgacattgt tgttaccaat
      781 cctccctaca agaacttaaa ggcggagaag ggccaatatt ctaatgactt ggaatatgaa
      841 atcgaccgtg ctcgctatgc tgaaattaag aagatggtca agcgcatctt caactactcc
      901 acagatggag tgttaaatct ttacaagctt ttcgtagaag aaattatcga taagtacgct
      961 aacccaaatg gttttgtatc attgttgatt ccgtcatcta tcttaacgga caaaacttgc
     1021 actaaattgc gtacccacat gttggtcgac tcaaacatcc tttcaattaa aatgattaac
     1081 gaaggtagcg gatacattga tgcccaacaa gctttgtccg cgattctgat tcagaaaggt
     1141 aagcgtacag aaagtatcaa ggtaaccaaa gattattcaa acaaccctaa ccagatcaca
     1201 gatatcaata tggaggacat tttgaatgaa aataccggca atgctatttt tgcgatcaat
     1261 aaccatgaat attttatttt aaagcagctt cgtaagtttc ctgtagtaaa agacttagac
     1321 ttcattatca atctgcgtgg tgagttggac cttacggcga acaaggatag tattgtcaac
     1381 attgacactg gctaccccct gcttcgtggc cgcaatatcg gctattatga aattctggac
     1441 acctgttccg gtgaatttgt atccaaggac tttatcgaaa actcaaagaa atcccgctat
     1501 atcaaagaga aacgcattgt atgtcagcag gtcgtgaaca tgaagaagga gcgccgcgtg
     1561 acctttgcgc tggtagagga gaactacgtt cttggaaact cctgtaattt tatcagcgta
     1621 atggacaatg actataacat cgatttatac gctattcttg gcctgttcaa cactagtatc
     1681 attaactggt tgtttaaact taccagctcg aataaccacg tgaataacta tgaaatcgac
     1741 tgtttccccg tgccaatcgg aagcccctat ctgaacaaga tttctaacct tgtgaagaaa
     1801 tatttgtcca acaaagattc atccttactt gaaaaaatcg aagagtatgc atacatcgcc
     1861 tatggaattc gtgaggcgaa ggaggataac gaagataagg acgatattgc taatctgaaa
     1921 gaaactaatg atattattaa gaagtattat tcagctatta aacacgtctt accgtcgatt
     1981 acacttgagg actcagtcag tattcttgag ggacaatcca gtattgagag ctttatcctt
     2041 cagtctggcg tagagttgga taaatacact cgtaatattg tgttgggtat cactgacaaa
     2101 tacatgaaga tcaagaaggg ggaaatcctt aatcatacga cgttcaaatt gagtgacctt
     2161 gatctggaaa tgatccgttc agtaccgccc gggggtaatt ggaaagatat ccctattgaa
     2221 actgtaaaga aatttaagcg tctgatgcgc atcacggaga cagggggacg cacaacactt
     2281 tatggccgca ttgactatga caagccgagt tataccatta cgacctactt taaccgccct
     2341 ggcaacggta cttatgtcca cccagttcac gatcgtgtct tgtcagtccg cgaggcggca
     2401 cgtttccagt gttttaagga tgattactat ttttacggaa acaagactca aatgcttaaa
     2461 caagtgggaa acgcagttcc aactattttg gcgtaccaga ttgcaaagaa aatcgttgac
     2521 aagacggggt gccgtaaatc gattgacttg ttctgtggag cagggggact gacagccggg
     2581 tttaaagaag cgggcattca gtcggtactg tgtaacgata ttgaggaatc ggcatgtatt
     2641 accctgaaga ttaataaccc ggagattaag gttttatgcg gagatattag ccaacacgaa
     2701 acaaaagaac acattgtaaa cgttgcaatc aacgaggatg tggacatcat ttgtggagga
     2761 ccgccatgtc aaggattttc catggctgga ctgcgtttga cggatgatcc ccgcaaccaa
     2821 cttttcaaag agtttatcga aatcgtaagt cgtgttaaac cgaaagtgat tgtctttgag
     2881 aatgtagaag ggattctttc cttccaatct ggcaaagtct accgtgcgat ccttgagatg
     2941 ttttccgaga ttgggtactt tacggaaggc cgtacgctta tgtctagcga ctatgcagtt
     3001 ccgcagaaac gcaagcgcgt ctttattatc tgcacgcgtg acgacatgga tgtaaaacct
     3061 gccgatttgt tccccacacc aattaccgag gagccagaat gccagattac agcacgtgac
     3121 acgattaaag atttggaaaa cattcaatgt gacgagaaag cgtgttacgt caaagttgag
     3181 cacgaaagcg acattcttaa agtatttaaa ggcaagatga cgtatcagga aatttactaa
     3241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaattc
     3661 agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg
     3721 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     3781 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     3841 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     3901 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     3961 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4141 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatcccacc ttgctcctgc
     4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4501 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4621 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaattg aaaaaggaag
     4801 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4861 cctgttatga catgcagata tttctatttc cgaaatagac agatcgctga gataggtgcc
     4921 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4981 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5041 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5221 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5461 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5641 cacctctgac ttgagcgtcg atttttgtga ttctcgtcag gggggcggag cctatggaaa
     5701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5761 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5821 gataccgctc gccgcagccg aacgaccaag cgcagcgagt cagtgagcga ggaagcggaa
     5881 g
//
