LOCUS       sequence-417255-        6213 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 6213)
  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 6213)
  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..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     315..333
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     351..370
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        351..369
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    369..393
                     /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)."
     terminator      2087..2133
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2503..3294,3295..3363))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMADHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3126..3145
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3364..3468)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3683..3700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3817..4362)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3897..3916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4470..4489)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4695..4772
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4773..5855
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4792..4811)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccctgaattc gcatctagat ggtagagcca caaacagccg gtacaagcaa cgatctccag
       61 gaccatctga atcatgcgcg gatgacacga actcacgacg gcgatcacag acattaaccc
      121 acagtacaga cactgcgaca acgtggcaat tcgtcgcaat accgtctcac tgaactggcc
      181 gataattgca gacgaacgtt attgtgtaat cgttaatccg caaataacgt aaaaacccgc
      241 ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcatc atgaccatga
      301 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcacctc taatacgact
      361 cactataggg aattgtgagc ggataacaat tccccagtac tgagcacact agttaacttt
      421 aagaaggaga catttgggga aaaaggccga atatggacag ggtcatccta tcttccttga
      481 gtacgctgaa cagatcattc aacacaagga gtaccagggt atgccagatc tgcgttatcc
      541 ggatgggcgt attcagtggg aggcaccttc taaccgtaaa tcgggcatct ttaaagacac
      601 caacatcaag cgtcgtaaat ggtgggagca aaaagcgatc tccattggaa tcgacccttc
      661 ttcgaatcag tggatctcca agacagcgaa attaatccac ccgacaatgc gtaaaccctg
      721 taagaagtgt ggacgtatta tggatcttcg ctactcgtat cccacaaaaa acctgatcaa
      781 gcgcatccgt aagttaccat atgtcgacga atcttttgaa atcgactcac tggagcatat
      841 tctgaagctg atcaaacgct tggtattaca atatggggac aaagtttatg acgatttacc
      901 caagctgtta acttgtaaag cggttaaaaa cattcctcgt ttgggaaatg atctggacac
      961 gtggttaaac tggattgact ccgtctatat ccctagcgaa ccatcaatgc tgtcaccggg
     1021 agctatggct aatccaccag atcgcttgga cgggttccac tcccttaatg agtgctgtcg
     1081 tagtcatgcg gatcgtggcc gctgggaaaa gaatcttcgc tcttatacaa ctgatcgtcg
     1141 cgcattcgaa tactgggtcg atggagactg ggtagcggct gataaattaa tgggacttat
     1201 ccgtaccaat gagcaaatca agaaggaaac atgtttaaac gataaccacc ctggtccttg
     1261 cagtgccgat catatcggtc cgatctctct gggatttgtc catcgtcctg aatttcaact
     1321 gctttgtaac tcctgtaatt ctgcaaagaa caaccgtatg actttcagcg acgttcagca
     1381 tcttatcaac gccgaaaata atggcgaaga ggtcgccagt tggtactgta aacatatctg
     1441 ggacttacgc aaacatgacg taaagaacaa tgaaaatgcg ttacgcctta gtaagatcct
     1501 tcgtgacaac cgtcacactg cgatgttcat tcttagtgag cttttgaaag acaatcatta
     1561 tctgttcctt tcaacgtttt taggccttca atatgcagag cgttcagtgt ccttttctaa
     1621 catcaagatt gaaaatcaca tcattactgg gcagatctcg gaacaacccc gtgacactaa
     1681 atatacagaa gaacaaaaag ctcgccgcat gcgcatcggc tttgaagccc ttaagagtta
     1741 catcgaaaag gagaaccgca acgccctttt ggtgatcaat gataagatca tcgacaaaat
     1801 caatgaaatc aagaacatcc ttcaggacat tcccgatgaa tacaagttat taaacgagaa
     1861 aatcagtgag caattcaata gcgaggaagt ctctgatgaa ttgttgcgtg atttggttac
     1921 acacctgcct acgaaggaat cagagccagc aaactttaag ctggcgcgca agtatttaca
     1981 agagatcatg gaaatcgtag gggacgaact gtccaagatg tgggaggacg aacgctatgt
     2041 tcgccagacc tttgccgatc tggattgata ataaatccag cagcattagc ataacccctt
     2101 ggggcctcta aacgggtctt gaggggtttt ttgaaaagat caaaggatct tctgtgatag
     2161 atactcgaac ccctagcccg ctcttatcgg gcggctaggg gttttttgtc caggtgctac
     2221 attgctgcca atgagacgac ggggtcatca cggctcatca tgcgccaaac aaatgtgtgc
     2281 aatacacgct cggatgactg catgatgacc gcactgactg gggacagcag atccacctaa
     2341 gcctgtgaga gaagcagaca cccgacagat caaggcagtt aactagtgca ctgcagtaca
     2401 gcggccgcga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2641 gcgggaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2881 atcaaggcga gttacatgat ccgccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgctc
     3181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttcctcg
     3481 aaaagtgcca cctgcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc
     3541 cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga tgagggtgtc
     3601 agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt
     3661 gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc gttcgactgc
     3721 ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc aggaagatac
     3781 ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc gcccccctga
     3841 caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg cctttcggtt
     3961 taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg acactcagtt
     4021 ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt cagtccgacc
     4081 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac
     4141 cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc atgcgccggt
     4201 taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca gttacctcgg
     4261 ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg
     4321 ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca tcttattaat
     4381 cagataaaat atttctagat ttcagtgcaa tttatctgcc ggacgcatcg tggccggcat
     4441 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
     4501 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
     4561 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
     4621 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
     4681 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     4741 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     4801 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     4861 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     4921 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     4981 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     5041 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5101 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5161 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5221 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5281 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5341 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5401 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5461 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     5521 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     5581 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     5641 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     5701 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     5761 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5821 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     5881 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
     5941 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     6001 atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt
     6061 cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc
     6121 gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc
     6181 gccggcatgc ttcaaatgta gcaccgcggc cgc
//
