LOCUS       sequence-12911-e        6657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6657)
  AUTHORS   Blumenthal A, Trujillo C, Ehrt S, Schnappinger D
  TITLE     Simultaneous analysis of multiple Mycobacterium tuberculosis 
            knockdown mutants in vitro and in vivo.
  JOURNAL   PLoS One. 2010 Dec 22;5(12):e15667. doi: 
            10.1371/journal.pone.0015667.
  PUBMED    21203517
REFERENCE   2  (bases 1 to 6657)
  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: "2010-12-22"; volume: "5(12):e15667. doi"; pages: " 
            10.1371/journal.pone.0015667"
COMMENT     Sequence Label: pGMCSq17-10M-0X
FEATURES             Location/Qualifiers
     source          1..6657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(174..220)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     477..499
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     496..512
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     496..512
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    532..552
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     606..622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     607..623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1561..1585
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1639..1656)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1809..1895
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1987..2014
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    complement(2041..2065)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2083..2103)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        2166..2184
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2166..2183
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2371..2390)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2372..2390)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2555..3143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2635..2654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3466..4257)
                     /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 gacgaggggc attcacacca gattggagct ggtgcagtga agagaataga ccgggacaag
       61 gttgcaccgg gagttgcagc ggtcggaacc ctcgccgtcg gcgggctggc gttcgccctg
      121 tcgttcacgg ctctagctga tcaccgcggc catgatggcc ggacaaacaa cagataaaac
      181 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gtcgatcgta
      241 cgctagttaa ctacgtcgac atcgatttaa ttaatatgca tcaattgatt taaatactag
      301 tgttaactat ttaattgggg accctagagg tccccttttt tattttaaaa attttttcna
      361 aggggacctc tagggtcccc agctggctag cgagtcatga ggtcaccatg ttctttcctg
      421 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccagcg
      481 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcta tcaactttgt
      541 atagaaaagt tgcatgacca acttcgataa cgttctcggc tcgatgatcc ccggcgggcc
      601 ccccctcgag gtcgacggta tcgatttaat taatatgcat caattgattt aaatactagt
      661 gttaactatt taattgggga ccctagaggt cccctttttt attttaaaaa ttttttcnaa
      721 ggggacctct agggtcccca gctggctagc gagtcatgag gtcaccatgg tcattaggag
      781 ccgctctcgc acttcagctg cttctccaag ccgcagatga tcagctccag gccgaacaag
      841 aaggccggct cggcgccctg gtggtcgaac agctcgatgg cctggcgcag caacggcggc
      901 atgctgtcgg tggtcggggt ctcgcgctcc tccttggcga cctggtgctc ctggtcctcc
      961 aacacgcagc ccagggtgaa gtggcccacg gccgacaggg cgtacaacgc gttctccagc
     1021 gagaagccct gctggcacag gaaggccaac tggttctcca gggtctcgta ctgcttctcg
     1081 gtcggccggg tgcccaggtg caccttggcg ccgtcgcggt ggctcagcaa ggcgcagcgg
     1141 aagctcttgg cgttgttgcg caggaagtcc tgccagctct cgccctccag cgggcagaag
     1201 tgggtgtggt ggcggtccaa catctcgatg gccagggcgt cgagcagggc ccgcttgttc
     1261 ttgacgtgcc agtacagggt cggctgctcg acgcccagct tctgggccag cttgcgggtg
     1321 gtcaggccct cgatgccgac ctcgttcagc agctccaggg cggagttgat gaccttggac
     1381 ttgtccaggc gggacattgc gaagtgattc ctcctgatgg taccggatcc gtggcaggag
     1441 ggtcgcctgc gtcggcaact cgcccaggct acacgagagc cctcgactgc gaaaatgccg
     1501 accaggcacc accgcactcc aggcttaacc cggtgcgacg atccaatatt acgagatctg
     1561 acaagtttgt acaaaaaagc aggctggctt aaccttacaa cgtcgaagct gttttggcgg
     1621 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     1681 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     1741 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     1801 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1861 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     1921 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1981 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtcg
     2041 tcacagcttt cttgtacaaa gtggttaatt aagcgcaagc ttcaacttta ttatacatag
     2101 ttgcttaagc tgcaggggac cctagaggtc ccctttttta ttttaaaaat tttttataac
     2161 cggtgattta ggtgacacta tagaagtgga attcgatggc ctagctggca tcggtacgtg
     2221 ccgcgcaacc atgtagtagt cctggagcgt gtccatctgg tgttcaagct taagaagtcc
     2281 aacacgccgc tcctccatcc gcaccaccct agttcgagtc ggaattcctg cgcaacggtc
     2341 aagcatgcca tatggaaacc gttgtggtct ccctatagtg agtcgtatta atttacgcgt
     2401 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2461 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     2761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3181 agtggaacta gcctaaaggt catccaccgg atcagcttag taaagccctc gctagatttt
     3241 aatgcggatg ttgcgattac ttcgccaact attgcgataa caagaaaaag ccagcctttc
     3301 atgatatatc tcccaatttg tgtagggctt attatgcacg cttaaaaata ataaaagcag
     3361 acttgacctg atagtttggc tgtgagcaat tatgtgctta gtgcatctaa cgcttgagtt
     3421 aagccgcgcc gcgaagcggc gtcggcttga acgaattgtt agacattatt tgccgactac
     3481 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     3541 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     3601 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     3661 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     3721 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     3781 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     3841 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     3901 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     3961 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     4021 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     4081 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     4141 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     4201 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatgat
     4261 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     4321 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     4381 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     4441 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     4501 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     4561 cccggcaacc ttgggcagca gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg
     4621 caaggtttcg gtctccacgc atcgtcaggc attggcggcc ttgctgttct tctacggcaa
     4681 ggtgctgtgc acggatctgc cctggcttca ggagatcgga agacctcggc cgtcgcggcg
     4741 cttgccggtg gtgctgaccc cggatgaagt ggttcgcatc ctcggttttc tggaaggcga
     4801 gcatcgtttg ttcgcccagc ttctgtatgg aacgggcatg cggatcagtg agggtttgca
     4861 actgcgggtc aaggatctgg atttcgatca cggcacgatc atcgtgcggg agggcaaggg
     4921 ctccaaggat cgggccttga tgttacccga gagcttggca cccagcctgc gcgagcaggg
     4981 gaattgatcc ggtggatgac cgctagctgg cgggaacggc aatgctcctg gcggcggcgg
     5041 ggctggtgcc caggtctccg cacagagcgg cggtgctggc gctcgcggcc aggcgtggtt
     5101 cttcgcgtac tgacaagaaa cccccctctt taggactcag tgtccttggg aggggggctt
     5161 tttgcgtttc aggaggtctt ggccagcttg gacatcgcct cagcgatagc ctcgtcgcgg
     5221 gcctcagacg ccatctggta cttcatcgcc atcctaggag tcgtgtgacc gagacgggcc
     5281 atcagctcct tggtcgtcgc acctgcctga gcggcgaacg tagcgccgac agcgcggagg
     5341 tcgtggatgc ggagttccgg ccgaccgatc ttggcgtagc cacgcttcag cgacttggtg
     5401 aacgcggact tcgacagccg gttgccctgc gtcgtggtca ccaggaatgc ctcggggccc
     5461 ttgttcatct tcgtacggtc cttcatgtgc gctcggatca tctccgcgac gtgaggcgga
     5521 accgtcacag gacgcttcga ccggacggtc ttggcgttgc caacgacgat cttgttcccc
     5581 acgcgggaag cgccacggcg cacccggagc ttcatcgtca tgccgtcgtc cacgatgtcc
     5641 ttgcggcgaa gctcgatcag ctctccgaac cggaggctcg tccacgccag gatgtatgcc
     5701 gcgatccggt agtgctcgaa gatctcagcg gcgacgatgt ccagctcctc aggcgtcagc
     5761 gcctctacgt cgcgctcatc ggctgccttc tgctcgatcc ggcacgggtt ctctgcgatc
     5821 agcttgtcct cgaccgctgt gttcatcacc gcccggagga cgttgtaggc atgccggcgg
     5881 gcagtcgggt gcttcctacc catcccggcc caccacgcac gcaccagagc tggcgtcatc
     5941 tctgtgaccg ccacttcacc tagcaccggg tagatgcggc gctccgcgtg cccgctgtac
     6001 agatccctgg tgccgtctgc gaggtcgcgc tccacgagcc acttccgggt gtactcctcc
     6061 agcgtgatgg cgctggcggc tgccttcttc gcccggtcct gtggaggggt ccaggtctcc
     6121 atctcgatga gccgcttctc gcccgcgagc caggcttcgg cgtccatctt gttgtcgtag
     6181 gtctgcagcg cgtagtacct cacaccgtcc tgcgggttga cgtatgaggc ttggatcctc
     6241 ccgctgcgct gagtcttcag cgatccccat ccgcgacgtg ccaactaggt ctcctctcgt
     6301 cgtgaacaag gctaccgggt tgcaactcct gtgcaactct caggcttcaa cgcgcttcta
     6361 cgacctgcaa tttctttcca cttagaggat gcagccgaga gggggtaaaa acctatcttg
     6421 accggcccat atatgtggtc ggcagacacc cattcttcca aactagctac gcgggttcga
     6481 ttcccgtcgc ccgctccgct ggtcagaggg tgttttcgcc ctctggccat ttttctttcc
     6541 aggggtctgc aactcttgtg cgactcttct gacctgggca tacgcggttg caacgcatcc
     6601 ctgatctggc tactttcgat gctgacaaac gaatagagcc ccccgcctgc gcgaaca
//
