LOCUS       sequence-12882-e        6108 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 6108)
  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 6108)
  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-MCKq20
FEATURES             Location/Qualifiers
     source          1..6108
                     /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(2365..2384)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2366..2384)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2467..3055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2547..2566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3387..4202)
                     /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     4110..4129
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4194..4213
                     /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 agcttgcaac ttcttcaaga
     2281 ggctgccgtc ccgcgccgat accatgctat agtatggcgt tcttgctgct gatggagcat
     2341 gccatatgga aaccgttgtg gtctccctat agtgagtcgt attaatttac gcgtttcact
     2401 ggccgacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2461 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2521 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2581 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2641 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2701 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2761 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2821 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2881 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     2941 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3001 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3061 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac tagtgaggtc tgcctcgtga
     3121 agaaggtgtt gctgactcat accaggcctg aatcgcccca tcatccagcc agaaagtgag
     3181 ggagccacgg ttgatgagag ctttgttgta ggtggaccag ttggtgattt tgaacttttg
     3241 ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct tcaactcagc
     3301 aaaagttcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat gctctgccag
     3361 tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     3421 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     3481 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     3541 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     3601 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag attatgcatt
     3661 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     3721 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     3781 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     3841 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     3901 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     3961 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     4021 acgctacctt tgccatgttt cagaaacaac tctggcgcat ggggcttccc atacaagcga
     4081 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4141 gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc
     4201 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     4261 tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtcgct ttgttgctag
     4321 ctggcgggaa cggcaatgct cctggcggcg gcggggctgg tgcccaggtc tccgcacaga
     4381 gcggcggtgc tggcgctcgc ggccaggcgt ggttcttcgc gtactgacaa gaaacccccc
     4441 tctttaggac tcagtgtcct tgggaggggg gctttttgcg tttcaggagg tcttggccag
     4501 cttggacatc gcctcagcga tagcctcgtc gcgggcctca gacgccatct ggtacttcat
     4561 cgccatccta ggagtcgtgt gaccgagacg ggccatcagc tccttggtcg tcgcacctgc
     4621 ctgagcggcg aacgtagcgc cgacagcgcg gaggtcgtgg atgcggagtt ccggccgacc
     4681 gatcttggcg tagccacgct tcagcgactt ggtgaacgcg gacttcgaca gccggttgcc
     4741 ctgcgtcgtg gtcaccagga atgcctcggg gcccttgttc atcttcgtac ggtccttcat
     4801 gtgcgctcgg atcatctccg cgacgtgagg cggaaccgtc acaggacgct tcgaccggac
     4861 ggtcttggcg ttgccaacga cgatcttgtt ccccacgcgg gaagcgccac ggcgcacccg
     4921 gagcttcatc gtcatgccgt cgtccacgat gtccttgcgg cgaagctcga tcagctctcc
     4981 gaaccggagg ctcgtccacg ccaggatgta tgccgcgatc cggtagtgct cgaagatctc
     5041 agcggcgacg atgtccagct cctcaggcgt cagcgcctct acgtcgcgct catcggctgc
     5101 cttctgctcg atccggcacg ggttctctgc gatcagcttg tcctcgaccg ctgtgttcat
     5161 caccgcccgg aggacgttgt aggcatgccg gcgggcagtc gggtgcttcc tacccatccc
     5221 ggcccaccac gcacgcacca gagctggcgt catctctgtg accgccactt cacctagcac
     5281 cgggtagatg cggcgctccg cgtgcccgct gtacagatcc ctggtgccgt ctgcgaggtc
     5341 gcgctccacg agccacttcc gggtgtactc ctccagcgtg atggcgctgg cggctgcctt
     5401 cttcgcccgg tcctgtggag gggtccaggt ctccatctcg atgagccgct tctcgcccgc
     5461 gagccaggct tcggcgtcca tcttgttgtc gtaggtctgc agcgcgtagt acctcacacc
     5521 gtcctgcggg ttgacgtatg aggcttggat cctcccgctg cgctgagtct tcagcgatcc
     5581 ccatccgcga cgtgccaact aggtctcctc tcgtcgtgaa caaggctacc gggttgcaac
     5641 tcctgtgcaa ctctcaggct tcaacgcgct tctacgacct gcaatttctt tccacttaga
     5701 ggatgcagcc gagagggggt aaaaacctat cttgaccggc ccatatatgt ggtcggcaga
     5761 cacccattct tccaaactag ctacgcgggt tcgattcccg tcgcccgctc cgctggtcag
     5821 agggtgtttt cgccctctgg ccatttttct ttccaggggt ctgcaactct tgtgcgactc
     5881 ttctgacctg ggcatacgcg gttgcaacgc atccctgatc tggctacttt cgatgctgac
     5941 aaacgaatag agccccccgc ctgcgcgaac agacgagggg cattcacacc agattggagc
     6001 tggtgcagtg aagagaatag accgggacaa ggttgcaccg ggagttgcag cggtcggaac
     6061 cctcgccgtc ggcgggctgg cgttcgccct gtcgttcacg gctctagc
//
