LOCUS       sequence-12913-e        2984 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2984)
  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 2984)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pEN12A-P1
FEATURES             Location/Qualifiers
     source          1..2984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(400..499)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        541..645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(646..714,715..1506)
                     /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     complement(864..883)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1677..2265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2166..2185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2419..2436
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2595..2622)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2714..2800)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2849..2871
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2863..2880
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2864..2880
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    join(2896..2984,1..11)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aaaagcaggc taccaggcct agatctgggg accctagagg tccccttttt tattttaaaa
       61 attttttcac gtgccagccg attggactag aaatattgga tcgtcggcac cgtcacggcc
      121 gtgggaggcg gcacgatccg cgacgtgatg atcggccgca tccccacggt gctgcgcagt
      181 gagctctacg ccatcccggc gttgatctgt gcgttcgcac gcacaggccc ggtgtgagaa
      241 gggtctctga cgagcgggag aactccctat cagtgataga gtttgtcctc cctatcagtg
      301 atagataggc tctgggagta cccgtgtgta cgaccagcac ggcatacatc atttcgacgc
      361 cgagagattc gccgcccgaa atgagcacga tccgcatgca cccagctttc ttgtacaaag
      421 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
      481 aaaataaaat cattatttgc catccagctg atgtcaggtg gcacttttcg gggaaatgtg
      541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      661 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      721 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      781 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      841 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      901 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      961 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1021 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1081 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1141 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1201 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1261 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1321 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1381 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1441 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1501 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1561 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1621 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1681 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1741 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1801 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1861 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1921 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1981 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2041 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2101 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2161 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2221 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2281 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2341 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2401 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2461 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2521 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     2581 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     2641 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     2701 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2761 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2821 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     2881 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     2941 cacattgatg agcaatgctt ttttataatg ccaactttgt acaa
//
