LOCUS       sequence-270631-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli 'blaTEM-1 with M. tuberculosis mpt63 signal sequence under 
            the control of hsp60 promoter that can be used as a reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  AUTHORS   McCann JR, McDonough JA, Pavelka MS, Braunstein M
  TITLE     Beta-lactamase can function as a reporter of bacterial protein 
            export during Mycobacterium tuberculosis infection of host cells.
  JOURNAL   Microbiology. 2007 Oct;153(Pt 10):3350-9. doi: 
            10.1099/mic.0.2007/008516-0.
  PUBMED    17906134
REFERENCE   2  (bases 1 to 5354)
  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: 
            "Microbiology."; date: "2007-10"; volume: "153(Pt 10):3350-9. doi"; 
            pages: " 10.1099/mic.0.2007/008516-0"
COMMENT     Sequence Label: pJES103
FEATURES             Location/Qualifiers
     source          1..5354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3084..3093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3137..3925
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(3283..3302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4002..4048
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4198..4217)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4209..5024
                     /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     complement(4282..4301)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaggg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcaacgcgtg agcccaccag ctccgtaagt tcgggcgctg
      781 tgtggctcgt acccgcgcat tcaggcggca gggggtctaa cgggtctaag gcggcgtgta
      841 cggccgccac agcggctctc agcggcccgg aaacgtcctc gaaacgacgc atgtgttcct
      901 cctggttggt acaggtggtt gggggtgctc ggctgtcgct ggtgttccac caccagggct
      961 cgacgggaga gcgggggagt gtgcagttgt ggggtggccc ctcagcgaaa tatctgactt
     1021 ggagctcgtg tcggaccata caccggtgat taatcgtggt ctactaccaa gcgtgagcca
     1081 cgtcgccgac gaatttgagc agctctggct gccgtactgg ccgctggcaa gcgacgatct
     1141 gctcgagggg atctaccgcc aaagccgcgc gtcggcccta ggccgccggt acatcgaggc
     1201 gaacccaaca gcgctggcaa acctgctggt cgtggacgta gaccatccag acgcagcgct
     1261 ccgagcgctc agcgcccggg ggtcccatcc gctgcccaac gcgatcgtgg gcaatcgcgc
     1321 caacggccac gcacacgcag tgtgggcact caacgcccct gttccacgca ccgaatacgc
     1381 gcggcgtaag ccgctcgcat acatggcggc gtgcgccgaa ggccttcggc gcgccgtcga
     1441 tggcgaccgc agttactcag gcctcatgac caaaaacccc ggccacatcg cctgggaaac
     1501 ggaatggctc cactcagatc tctacacact cagccacatc gaggccgagc tcggcgcgaa
     1561 catgccaccg ccgcgctggc gtcagcagac cacgtacaaa gcggctccga cgccgctagg
     1621 gcggaattgc gcactgttcg attccgtcag gttgtgggcc tatcgtcccg ccctcatgcg
     1681 gatctacctg ccgacccgga acgtggacgg actcggccgc gcgatctatg ccgagtgcca
     1741 cgcgcgaaac gccgaatttc cgtgcaacga cgtgtgtccc ggaccgctac cggacagcga
     1801 ggtccgcgcc atcgccaaca gcatttggcg ttggatcaca accaagtcgc gcatttgggc
     1861 ggacgggatc gtggtctacg aggccacact cagtgcgcgc cagtcggcca tctcgcggaa
     1921 gggcgcagca gcgcgcacgg cggcgagcac agttgcgcgg cgcgcaaagt ccgcgtcagc
     1981 catggaggca ttgctatgag cgacggctac agcgacggct acaaccggca gccgactgtc
     2041 cgcaaaaagc ggcgcgtgac cgccgccgaa ggcgctcgaa tcaccggact atccgaacgc
     2101 cacgtcgtcc ggctcgtggc gcaggaacgc agcgagtggc tcgccgagca ggctgcacgc
     2161 cgcgaacgca tccgcgccta tcacgacgac gagggccact cttggccgca aacggccaaa
     2221 catttcgggc tgcatctgga caccgttaag cgactcggct atcgggcgag gaaagagcgt
     2281 gcggcagaac aggaagcggc tcaaaaggcc cacaacgaag ccgacaatcc accgctgttc
     2341 taacgcaatt ggggagcggg tgtcgcgggg gttccgtggg gggttccgtt gcaacgggtc
     2401 ggacaggtaa aagtcctggt agacgctagt tttctggttt gggccatgcc tgtctcgttg
     2461 cgtgtttcgt tgcgcccgtt ttgaatacca gccagacgag acggggttct acgaatcttg
     2521 gtcgatacca agccatttcc gctgaatatc ggggagctca ccgccagaat cggtggttgt
     2581 ggtgatgtac gtggcgaact ccgttgtagt gcctgtggtg gcatccgtgg cgcggccgcg
     2641 gtaccagatc tttaaatcta gacggtgacc acaacgcgcc cgctttgatc ggggacgtct
     2701 gcggccgacc atttacgggt cttgttgtcg ttggcggtca tgggccgaac atactcaccc
     2761 ggatcggagg gccgaggaca aggtcgaacg aggggcatga cccggtgcgg ggcttcttgc
     2821 actcggcata ggcgagtgct aagaataacg ttggcactcg cgaccggtga gtgctaggtc
     2881 gggacggtga ggccaggccc gtcgtcgcag cgagtggcag cgaggacaac ttgagccgtc
     2941 cgtcgcgggc actgcgcccg gccagcgtaa gtagcggggt tgccgtcacc cggtgacccc
     3001 cgtttcatcc ccgatccgga ggaatcactt cgcaatgggc atgaagctca ccacaatgat
     3061 caagacggca gtagcggtcg tggccatggc ggccatcgcg acctttgcgg caccggtcgc
     3121 gttggctgga tctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3181 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3241 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3301 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3361 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3421 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3481 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3541 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3601 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3661 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3721 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3781 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3841 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3901 tgcctcactg attaagcatt ggtaacgtgg gatccagctg cagaattcga agcttatcga
     3961 tgtcgacgta gttaactagc gtacgatcga ctgccaggca tcaaataaaa cgaaaggctc
     4021 agtcgaaaga ctgggccttt cgttttatgc catcatggcc gcggtgatca gctagccacc
     4081 tgacgtcggg ggggggggaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     4141 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     4201 ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga ttaaattcca
     4261 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     4321 cgacaatcta tcgcttgtat gggaagcccc atgcgccaga gttgtttctg aaacatggca
     4381 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     4441 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     4501 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     4561 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     4621 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     4681 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     4741 tctggaaaga aatgcataat cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     4801 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     4861 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     4921 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     4981 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     5041 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt gttgaataaa
     5101 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     5161 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcaccaa
     5221 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
     5281 aacaccttct tcacgaggca gacctcacta gttccatgag cgtcagaccc cgtagaaaag
     5341 atcaaaggat cttc
//
