LOCUS       sequence-69380-e        4567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for ectopic hypoxia inducible expression of mycobacterial proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4567)
  AUTHORS   Parikh A, Kumar D, Chawla Y, Kurthkoti K, Khan S, Varshney U, 
            Nandicoori VK
  TITLE     Development of a new generation of vectors for gene expression, gene
            replacement, and protein-protein interaction studies in 
            mycobacteria.
  JOURNAL   Appl Environ Microbiol. 2013 Mar;79(5):1718-29. doi: 
            10.1128/AEM.03695-12. Epub 2013 Jan 11.
  PUBMED    23315736
REFERENCE   2  (bases 1 to 4567)
  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"; doi: 
            "10.1128/AEM.03695-12"; journalName: "Appl Environ Microbiol"; date:
            "2013-03"; volume: "79"; issue: "5"; pages: "1718-29"
COMMENT     Sequence Label: pST-KNarK2
FEATURES             Location/Qualifiers
     source          1..4567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             337..354
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             427..450
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      complement(469..511)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      540..586
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(736..755)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             747..1562
                     /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(820..839)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1894..2482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2383..2402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agtactatct ggtcgccgca catgatgttg ctcccctttc cagtggcgac caggctcagc
       61 tcgctggtca taatgcatcc ttgcgattgg ggtgtaacct ccacagacaa atccgaccac
      121 cgcgccggag ccctccgata gggcctgaag tcccggccac tggggtcatt cgtccgccag
      181 gggcgaaagg tctgccacat tggggacttc cggccctaac tgagccggcg ccatcgaagc
      241 cagggttagc gcatggctgt ccgggcgggg cgcggagttc gccggagtcc gagaccccgg
      301 atcgtgtcgc gcatgcggag gaatcacttc gcaatgcacc accaccacca ccatatggga
      361 tccgatatct ctagagcggc cgcacagctg gggcccgaat tcctgcagca cgtgttaatt
      421 aagcttgact acaaggacga cgacgacaag taaaagctat cagttaacta aaacgaaagg
      481 ctcagtcgaa agactgggcc tttcgtttta taactagcgt acgatcgact gccaggcatc
      541 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatgcca tcatggccgc
      601 ggtgatcagc tagccacctg acgtcggggg ggggggaaag ccacgttgtg tctcaaaatc
      661 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
      721 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcga
      781 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
      841 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagccccat gcgccagagt
      901 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
      961 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     1021 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
     1081 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     1141 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     1201 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     1261 gctggcctgt tgaacaagtc tggaaagaaa tgcataatct tttgccattc tcaccggatt
     1321 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     1381 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     1441 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     1501 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     1561 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     1621 gcggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca cgcatcttcc
     1681 cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact ggtccaccta
     1741 caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg gggcgattca
     1801 ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctcactagt tccactgagc
     1861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2041 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2281 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     2461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2581 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcaacgc
     2641 gttacgtaga gcccaccagc tccgtaagtt cgggtgctgt gtggctcgta cccgcgcatt
     2701 caggcggcag ggggtctaac gggtctaagg cggcgtgtac ggccgccaca gcggctctta
     2761 gcggcccgga aacgtcctcg aaacgacgca tgtgttcctc ctggttggta caggtggttg
     2821 ggggtgctcg gctgtcgctg gtgtttcatc atcagggctc gacgggagag cgggggagtg
     2881 tgcagttgtg gggtggcccc tcagcgaaat atctgacttg gagctcgtgt cggaccatac
     2941 accggtgatt aatcgtggtt tattatcaag cgtgagccac gtcgccgacg aatttgagca
     3001 gctctggctg ccgtactggt ccctggcaag cgacgatctg ctcgagggga tctaccgcca
     3061 aagccgcgcg tcggccctag gccgccggta catcgaggcg aacccaacag cgctggcaaa
     3121 cctgctggtc gtggacgtag accatccaga cgcagcgctc cgagcgctca gcgcccgggg
     3181 gtcccatccg ctgcccaacg cgatcgtggg caatcgcgcc aacggccacg cacacgcagt
     3241 gtgggcactc aacgcccctg ttccacgcac cgaatacgcg cggcgtaagc cgctcgcata
     3301 catggcggcg tgcgccgaag gccttcggcg cgccgtcgat ggcgaccgca gttactcagg
     3361 cctcatgacc aaaaaccccg gccacatcgc ctgggaaacg gaatggctcc actcagatct
     3421 ctacacactc agccacatcg aggccgagct cggcgcgaac atgccaccgc cgcgctggcg
     3481 tcagcagacc acgtacaaag cggctccgac gccgctaggg cggaattgcg cactgttcga
     3541 ttccgtcagg ttgtgggcct atcttcccgc cctcatgcgg atctacctgc cgacccggaa
     3601 cgtggacgga ctcggccgcg cgatctatgc cgagtgccac gcgcgaaacg ccgaatttcc
     3661 gtgcaacgac gtgtgtcccg gaccgctacc ggacagcgag gtccgcgcca tcgccaacag
     3721 catttggcgt tggatcacaa ccaagtcgcg catttgggcg gacgggatcg tggtctacga
     3781 ggccacactc agtgcgcgcc atgcggccat ctcgcggaag ggcgcagcag cgcgcacggc
     3841 ggcgagcaca gttgcgcggc gcgcaaagtc cgcgtcagcc atggaggcat tgctatgagc
     3901 gacggctaca gcgacggcta cagcgacggc tacaactggc agccgactgt ccgcaaaaag
     3961 cggcgcgtga ccgccgccga aggcgctcga atcaccggac tatccgaacg ccacgtcgtc
     4021 cggctcgtgg cgcaggaacg cagcgagtgg ttcgccgagc aggctgcacg ccgcgaacgc
     4081 atccgcgcct atcacgacga cgagggccac tcttggccgc aaacggccaa acatttcggg
     4141 ctgcatctgg acaccgttaa gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa
     4201 caggaagcgg ctcaaaaggc ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat
     4261 tggggagcgg gtgtcgcggg ggttccgtgg ggggttccgt tgcaacgggt cggacaggta
     4321 aaagtcctgg tagacgctag ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg
     4381 ttgcgtccgt tttgaatacc agccagacga gacggggttc tacgaatctt ggtcgatacc
     4441 aagccatttc cgctgaatat cgtggagctc accgccagaa tcggtggttg tggtgatgta
     4501 cgtggcgaac tccgttgtag tgcttgtggt ggcatccgtg gcggtaccgc ctgacgcccc
     4561 ccggatc
//
