LOCUS       sequence-343794-        2817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP expression under the control of E. coli dnaK promoter engineered
            with double IR2 HAIR motif from M. tuberculosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2817)
  AUTHORS   Saltepe B, Bozkurt EU, Haciosmanoglu N, Seker UOS
  TITLE     Genetic Circuits To Detect Nanomaterial Triggered Toxicity through 
            Engineered Heat Shock Response Mechanism.
  JOURNAL   ACS Synth Biol. 2019 Oct 18;8(10):2404-2417. doi: 
            10.1021/acssynbio.9b00291. Epub 2019 Oct 2.
  PUBMED    31536326
REFERENCE   2  (bases 1 to 2817)
  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: 
            "10.1021/acssynbio.9b00291"; journalName: "ACS Synth Biol"; date: 
            "2019-10-18- 18"; volume: "8"; issue: "10"; pages: "2404-2417"
COMMENT     Sequence Label: PdnaK-IR2-IR2 GFP pZa
FEATURES             Location/Qualifiers
     source          1..2817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     326..346
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      407..493
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(528..545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(662..1207)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(742..761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1321..1415)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1436..2095)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2010..2029
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2096..2198)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             2466..2477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2490..2817,1..386)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2520..2548)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt
       61 ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg
      121 tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca
      181 acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg
      241 atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc ctttcgaaag
      301 atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta
      361 cacatggcat ggatgaacta tacaaataaa agcttacgcg tggcatcaaa taaaacgaaa
      421 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      481 gagtaggaca aatccgccgc cctagaccta gggatatatt ccgcttcctc gctcactgac
      541 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
      601 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
      661 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
      721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
      781 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
      841 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
      901 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      961 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     1021 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     1081 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     1141 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     1201 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     1261 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgttac tagtgcttgg
     1321 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     1381 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
     1441 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     1501 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     1561 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     1621 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     1681 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     1741 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     1801 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     1861 ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     1921 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     1981 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2041 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2101 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2161 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccagatat
     2221 cgacgtcctc gagaaaagca caaaaaattt ttgcatctcc cccttgatga cgtggtttac
     2281 gaccccattt agtagtcaac cgcagtgagt gagtctgcaa aaaaatgaaa ttgggcagtt
     2341 gaaaccagac gtttcgcccc tattacagac tcacaaccac atgatgaccg aatatatagt
     2401 ggagacgttt agatgagtaa gttgagtgca tcaggctcag agtaagttga gtgcatcagg
     2461 ctcagaaaga ggagaaaacg cgtggtacca tgcgtaaagg agaagaactt ttcactggag
     2521 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     2581 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     2641 gaaaactacc tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct
     2701 ttgcgagata cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag
     2761 gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag acacgtg
//
