LOCUS       sequence-12857-e        6107 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 6107)
  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 6107)
  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: pDE43-MCKq19
FEATURES             Location/Qualifiers
     source          1..6107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(37..83)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     340..362
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     359..375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     359..375
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    395..518
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(559..864)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(661..680)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(1206..1865)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1780..1799
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(1919..1925,1926..1943,1944..1949))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(1974..2097)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2160..2178
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2160..2177
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2364..2383)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2365..2383)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2466..3054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2546..2565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3386..4201)
                     /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
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4109..4128
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4193..4212
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgatcaccgc ggccatgatg gccggacaaa caacagataa aacgaaaggc ccagtctttc
       61 gactgagcct ttcgttttat ttgatgcctg gcagtcgatc gtacgctagt taactacgtc
      121 gacatcgatt taattaatat gcatcaattg atttaaatac tagtgttaac tatttaattg
      181 gggaccctag aggtcccctt ttttatttta aaaatttttt cnaaggggac ctctagggtc
      241 cccagctggc tagcgagtca tgaggtnacc atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgatacca gcggataaca atttcacaca
      361 ggaaacagct atgaccatga ttacgccaag ctatcaactt tgtatagaaa agttgaacga
      421 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
      481 acataatact gtaaaacaca acatatccag tcactatggt cgacctgcag actggctgtg
      541 tataagggag cctgacattt atattcccca gaacatcagg ttaatggcgt ttttgatgtc
      601 attttcgcgg tggctgagat cagccacttc ttccccgata acggagaccg gcacactggc
      661 catatcggtg gtcatcatgc gccagctttc atccccgata tgcaccaccg ggtaaagttc
      721 acgggggact ttatctgaca gcagacgtgc actggccagg gggatcacca tccgtcgccc
      781 gggcgtgtca ataatatcac tctgtacatc cacaaacaga cgataacggc tctctctttt
      841 ataggtgtaa accttaaact gcatttcacc agcccctgtt ctcgtcggca aaagagccgt
      901 tcatttcaat aaaccgggcg acctcagcca tcccttcctg attttccgct ttccagcgtt
      961 cggcacgcag acgacgggct tcattctgca tggttgtgct taccgaaccg gagatattga
     1021 catcatatat gccttgagca actgatagct gtcgctgtca actgtcactg taatacgctg
     1081 cttcatagca tacctctttt tgacatactt cgggtataca tatcagtata tattcttata
     1141 ccgcaaaaat cagcgcgcaa atacgcatac tgttatctgg cttttagtaa gccggatcct
     1201 ctagattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     1261 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1321 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     1381 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     1441 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     1501 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     1561 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     1621 agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga
     1681 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     1741 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     1801 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     1861 tccattttag cttccttagc tcctgaaaat ctcgacggat cctaactcaa aatccacaca
     1921 ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgcggc cgccatagtg
     1981 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     2041 atatattgat atttatatca ttttacgttt ctcgttcaac tttattatac atagttgctt
     2101 aagctgcagg ggaccctaga ggtccccttt tttattttaa aaatttttta taaccggtga
     2161 tttaggtgac actatagaag tggaattcga tggcctagct ggcatcggta cgtgccgcgc
     2221 aaccatgtag tagtcctgga gcgtgtccat ctggtgttca agctttcggc tagatgtgca
     2281 acgcatggca gtcgccaatc cacggccgaa gcagagcgct atttcactga cgcgtgcatg
     2341 ccatatggaa accgttgtgg tctccctata gtgagtcgta ttaatttacg cgtttcactg
     2401 gccgacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2641 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2881 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3061 cctttgatct tttctacggg gtctgacgct cagtggaact agtgaggtct gcctcgtgaa
     3121 gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg
     3181 gagccacggt tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc
     3241 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca
     3301 aaagttcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt
     3361 gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca
     3421 atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag
     3481 gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc
     3541 cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa
     3601 gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaaga ttatgcattt
     3661 ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa
     3721 ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa
     3781 aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa
     3841 caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga
     3901 tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa
     3961 gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa
     4021 cgctaccttt gccatgtttc agaaacaact ctggcgcatg gggcttccca tacaagcgat
     4081 agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag
     4141 catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca
     4201 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     4261 ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtcgctt tgttgctagc
     4321 tggcgggaac ggcaatgctc ctggcggcgg cggggctggt gcccaggtct ccgcacagag
     4381 cggcggtgct ggcgctcgcg gccaggcgtg gttcttcgcg tactgacaag aaacccccct
     4441 ctttaggact cagtgtcctt gggagggggg ctttttgcgt ttcaggaggt cttggccagc
     4501 ttggacatcg cctcagcgat agcctcgtcg cgggcctcag acgccatctg gtacttcatc
     4561 gccatcctag gagtcgtgtg accgagacgg gccatcagct ccttggtcgt cgcacctgcc
     4621 tgagcggcga acgtagcgcc gacagcgcgg aggtcgtgga tgcggagttc cggccgaccg
     4681 atcttggcgt agccacgctt cagcgacttg gtgaacgcgg acttcgacag ccggttgccc
     4741 tgcgtcgtgg tcaccaggaa tgcctcgggg cccttgttca tcttcgtacg gtccttcatg
     4801 tgcgctcgga tcatctccgc gacgtgaggc ggaaccgtca caggacgctt cgaccggacg
     4861 gtcttggcgt tgccaacgac gatcttgttc cccacgcggg aagcgccacg gcgcacccgg
     4921 agcttcatcg tcatgccgtc gtccacgatg tccttgcggc gaagctcgat cagctctccg
     4981 aaccggaggc tcgtccacgc caggatgtat gccgcgatcc ggtagtgctc gaagatctca
     5041 gcggcgacga tgtccagctc ctcaggcgtc agcgcctcta cgtcgcgctc atcggctgcc
     5101 ttctgctcga tccggcacgg gttctctgcg atcagcttgt cctcgaccgc tgtgttcatc
     5161 accgcccgga ggacgttgta ggcatgccgg cgggcagtcg ggtgcttcct acccatcccg
     5221 gcccaccacg cacgcaccag agctggcgtc atctctgtga ccgccacttc acctagcacc
     5281 gggtagatgc ggcgctccgc gtgcccgctg tacagatccc tggtgccgtc tgcgaggtcg
     5341 cgctccacga gccacttccg ggtgtactcc tccagcgtga tggcgctggc ggctgccttc
     5401 ttcgcccggt cctgtggagg ggtccaggtc tccatctcga tgagccgctt ctcgcccgcg
     5461 agccaggctt cggcgtccat cttgttgtcg taggtctgca gcgcgtagta cctcacaccg
     5521 tcctgcgggt tgacgtatga ggcttggatc ctcccgctgc gctgagtctt cagcgatccc
     5581 catccgcgac gtgccaacta ggtctcctct cgtcgtgaac aaggctaccg ggttgcaact
     5641 cctgtgcaac tctcaggctt caacgcgctt ctacgacctg caatttcttt ccacttagag
     5701 gatgcagccg agagggggta aaaacctatc ttgaccggcc catatatgtg gtcggcagac
     5761 acccattctt ccaaactagc tacgcgggtt cgattcccgt cgcccgctcc gctggtcaga
     5821 gggtgttttc gccctctggc catttttctt tccaggggtc tgcaactctt gtgcgactct
     5881 tctgacctgg gcatacgcgg ttgcaacgca tccctgatct ggctactttc gatgctgaca
     5941 aacgaataga gccccccgcc tgcgcgaaca gacgaggggc attcacacca gattggagct
     6001 ggtgcagtga agagaataga ccgggacaag gttgcaccgg gagttgcagc ggtcggaacc
     6061 ctcgccgtcg gcgggctggc gttcgccctg tcgttcacgg ctctagc
//
