LOCUS       sequence-12914-e        4825 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 4825)
  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 4825)
  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: pDO23A
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(141..160)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(820..1611,1612..1680))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1443..1462
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1681..1785)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1847..2078
                     /gene="mutant version of attP"
                     /label=attP3
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2435..2740)
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2537..2556)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(3082..3741)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3656..3675
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3742..3844)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    4002..4233
                     /gene="mutant version of attP"
                     /label=attP2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4271..4288)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4271..4287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4280..4302)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      4351..4437
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4529..4556
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4715..4732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
       61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      901 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1141 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1441 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1681 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1741 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1801 agtgccacct gacatcgcgg ccgcgttaac gctaccatgg agctccaaat aatgatttta
     1861 ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt ttttataatg
     1921 ccaactttgt ataataaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
     1981 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca
     2041 ctatgaatca actacttaga tggtattagt gacctgtaga attcgagctc gaccgacagc
     2101 cttccaaatg ttcttctcaa acggaatcgt cgtatccagc ctactcgcta ttgtcctcaa
     2161 tgccgtatta aatcataaaa agaaataaga aaaagaggtg cgagcctctt ttttgtgtga
     2221 caaaataaaa acatctacct attcatatac gctagtgtca tagtcctgaa aatcatctgc
     2281 atcaagaaca atttcacaac tcttatactt ttctcttaca agtcgttcgg cttcatctgg
     2341 attttcagcc tctatactta ctaaacgtga taaagtttct gtaatttcta ctgtatcgac
     2401 ctgcagactg gctgtgtata agggagcctg acatttatat tccccagaac atcaggttaa
     2461 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
     2521 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
     2581 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
     2641 tcaccatccg tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat
     2701 aacggctctc tcttttatag gtgtaaacct taaactgcat ttcaccagtc cctgttctcg
     2761 tcagcaaaag agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt
     2821 tccgctttcc agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc
     2881 agaccggaga tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg
     2941 tcactgtaat acgctgcttc atagcacacc tctttttgac atacttcggg tatacatatc
     3001 agtatatatt cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt
     3061 tagtaagccg gatccacgcg attacgcccc gccctgccac tcatcgcagt actgttgtaa
     3121 ttcattaagc attctgccga catggaagcc atcacagacg gcatgatgaa cctgaatcgc
     3181 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     3241 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     3301 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     3361 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     3421 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     3481 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
     3541 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     3601 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     3661 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     3721 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     3781 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     3841 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
     3901 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
     3961 gtcgggtgat gctgccaact tagtcgacgc tctagagctc caaataatga ttttattttg
     4021 actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttttta taatgccaac
     4081 tttgtacaag aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta
     4141 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatgc agtcactatg
     4201 aaccaactac ttagatggta ttagtgacct gtagaattcg agctctagag ctgcagggcc
     4261 cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg
     4321 ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac
     4381 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4441 ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4501 ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg
     4561 tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg
     4621 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4681 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4801 aatcagggga taacgcagga aagaa
//
