LOCUS       sequence-270627-        5282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli 'blaTEM-1 (without signal sequence) that can be used as a 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5282)
  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 5282)
  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: pJES102
FEATURES             Location/Qualifiers
     source          1..5282
                     /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"
     CDS             3047..3835
                     /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(3193..3212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3930..3976
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4126..4145)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4137..4952
                     /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(4210..4229)
                     /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 cgcaatggga tctcacccag aaacgctggt
     3061 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     3121 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     3181 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     3241 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     3301 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     3361 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     3421 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3481 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3541 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3601 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3661 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3721 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3781 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaacgtgg
     3841 gatcccaaga caattgcgga tccagctgca gaattcgaag cttatcgatg tcgacgtagt
     3901 taactagcgt acgatcgact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     3961 gggcctttcg ttttatgcca tcatggccgc ggtgatcagc tagccacctg acgtcggggg
     4021 ggggggaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     4081 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     4141 gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg
     4201 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     4261 gcttgtatgg gaagccccat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     4321 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     4381 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     4441 ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     4501 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     4561 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     4621 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     4681 tgcataatct tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     4741 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     4801 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     4861 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     4921 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     4981 ggcagagcat tacgctgact tgacgggacg gcggctttgt tgaataaatc gaacttttgc
     5041 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     5101 aaagttcaaa atcaccaact ggtccaccta caacaaagct ctcaccaacc gtggctccct
     5161 cactttctgg ctggatgatg gggcgattca ggcctggtat gagtcagcaa caccttcttc
     5221 acgaggcaga cctcactagt tccatgagcg tcagaccccg tagaaaagat caaaggatct
     5281 tc
//
