LOCUS       sequence-342221-        2791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP expression under the control of E. coli dnaK promoter engineered
            with IR2 HAIR motif from M. tuberculosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2791)
  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 2791)
  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 GFP pZa
FEATURES             Location/Qualifiers
     source          1..2791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      38..124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(159..176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(293..838)
                     /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(373..392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(952..1046)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1067..1726)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1641..1660
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1727..1829)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             2071..2082
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2095..2791,1..17)
                     /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(2125..2153)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2748..2768
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 tggatgaact atacaaataa aagcttacgc gtggcatcaa ataaaacgaa aggctcagtc
       61 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      121 aaatccgccg ccctagacct agggatatat tccgcttcct cgctcactga ctcgctacgc
      181 tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga tttcctggaa
      241 gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg tttttccata
      301 ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc
      361 cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg cgctctcctg
      421 ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca
      481 cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc
      541 cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa
      601 agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt
      661 gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc
      721 aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg
      781 caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga
      841 agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta tctcttcaaa
      901 tgtagcacct gaagtcagcc ccatacgata taagttgtta ctagtgcttg gattctcacc
      961 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     1021 cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct
     1081 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     1141 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     1201 atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     1261 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     1321 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     1381 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     1441 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     1501 ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     1561 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     1621 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     1681 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     1741 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     1801 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccagata tcgacgtcct
     1861 cgagaaaagc acaaaaaatt tttgcatctc ccccttgatg acgtggttta cgaccccatt
     1921 tagtagtcaa ccgcagtgag tgagtctgca aaaaaatgaa attgggcagt tgaaaccaga
     1981 cgtttcgccc ctattacaga ctcacaacca catgatgacc gaatatatag tggagacgtt
     2041 tagatgagta agttgagtgc atcaggctcg aaagaggaga aaacgcgtgg taccatgcgt
     2101 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     2161 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     2221 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     2281 actttcggtt atggtgttca atgctttgcg agatacccag atcatatgaa acagcatgac
     2341 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     2401 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     2461 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     2521 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     2581 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     2641 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     2701 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     2761 tttgtaacag ctgctgggat tacacatggc a
//
