LOCUS       sequence-73916-e        3880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ampicillin resistant; entry clone encoding reverse TetR (tetR38) and
            contains attL4 and attL1 sites for recombination into a Gateway 
            destination vector at the attR4 and attR1 sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3880)
  AUTHORS   Kim JH, O'Brien KM, Sharma R, Boshoff HI, Rehren G, Chakraborty S, 
            Wallach JB, Monteleone M, Wilson DJ, Aldrich CC, Barry CE 3rd, Rhee 
            KY, Ehrt S, Schnappinger D
  TITLE     A genetic strategy to identify targets for the development of drugs 
            that prevent bacterial persistence.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Nov 4.
  PUBMED    24191058
REFERENCE   2  (bases 1 to 3880)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Nov 4."
COMMENT     Sequence Label: pEN41A-T38S38
FEATURES             Location/Qualifiers
     source          1..3880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..40
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     32..49
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     33..49
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    89..184
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1370..1493
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1590..1694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1695..1763,1764..2555)
                     /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(1913..1932)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2726..3314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3215..3234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3468..3485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3644..3671)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3763..3849)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg
       61 gcccgagtta acgctaccat ggagctccaa ataatgattt tattttgact gatagtgacc
      121 tgttcgttgc aacaaattga taagcaatgc ttttttataa tgccaacttt gtatagaaaa
      181 gttgcagctg gctagcgagt catgaggtca ccatggtcat taggagccgc tctcgcactt
      241 cagctgcttc tccaagccgc agatgatcag ctccaggccg aacaagaagg ccggctcggc
      301 gccctggtgg tcgaacagct cgatggcctg gcgcagcaac ggcggcatgc tgtcggtggt
      361 cggggtctcg cgctcctcct tggcggccgt gtgctcctgg tcctccaaca cgcagcccag
      421 ggtgaagtgg cccacggccg acagggcgta caacgcgttc tccagcgaga agccgttctc
      481 ggtcatgaag cgcagctggg tctccacggt gtcgtactgc ttctcgtccg ggcgggtgcc
      541 caggtgcacc ttggcgccgt cgcggtagcg cagcagcgcg cggcggaacg acatggcgtt
      601 gttgcgcagg aacgactgcc acgactcgcc ggccgccggc agcgagtagt cgtggtggcg
      661 cgccaggatc tccaccgcca gggcgtccag cagggcccgc ttgttcttga cgtgccagta
      721 cagggtcttc tgcgcgacgc ccagcttctg ggccagcttg cgggtggtca cgccctcgat
      781 gccgacctcg ttgcccaggg ccagggcgga gttgatgacc ttggacttgt ccaggcggga
      841 cattgcgaag tgattcctcc tgatggtacc ctaggtcaga tcaacggcct ggtcgacaag
      901 ggcaatacgg tgatcgtcat cgaacataac ctggacgtga tcggccgtca actcgttgag
      961 gtaccggtcg gattcaaatg gttcgtcgac ggtctgatcc cgcggtgtgc accgatgccc
     1021 tcacggttcc agcggtcgac caggtcctcg ccgtgacgcc ccttgatcgc aaacgccacc
     1081 gaatgcacca cggccacagc caaagccgac acgctggtta ccacgacgat gttcacgggg
     1141 ggccctcgcg gatctggccc aactgctgga cttccaccga agcaatcgag ccgatgtcac
     1201 gctgcaactg gtgcgggtgg gcgacccgcg ggcattcggc tgcgtaccca ccgacgagga
     1261 ggaccgcgta gtcgcctttc tggagaagac ggagcagacg gtgctgttgg gcgcctgctg
     1321 gattcccagg tcttcgaaca ccgacacgct cgcggcgtga tccaatattc aagtttgtac
     1381 aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
     1441 cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact atgaaccaac
     1501 tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg gcggccgcga
     1561 tgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1621 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1681 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1741 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1801 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1861 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1921 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1981 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2041 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2101 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2161 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2221 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     2281 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2341 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2401 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2461 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2521 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2581 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2641 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3361 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3421 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3481 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3541 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3601 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3661 gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc
     3721 tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag
     3781 agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg
     3841 ttttatttga tgcctggcag ttccctactc tcgcgttaac
//
