LOCUS       sequence-343075-        2614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP expression under the control of E. coli clpB promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2614)
  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 2614)
  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: PclpB GFP pZa
FEATURES             Location/Qualifiers
     source          1..2614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6..92
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(127..144)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(261..806)
                     /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(341..360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(920..1014)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1035..1694)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1609..1628
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1695..1797)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             1868..1879
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1886..2599
                     /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(1916..1944)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2539..2559
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
       61 tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc ctagacctag ggatatattc
      121 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
      181 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
      241 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
      301 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      361 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
      421 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
      481 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
      541 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
      601 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
      661 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
      721 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
      781 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
      841 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
      901 agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc
      961 tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg
     1021 agctcgatat caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta
     1081 agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc
     1141 atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag
     1201 ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag
     1261 acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac
     1321 gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag
     1381 agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc
     1441 catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg
     1501 cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt
     1561 aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga
     1621 aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg
     1681 atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1741 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg
     1801 tctcattttc gccagatatc gacgtcctcg agcttgaata attgagggat gacctcattt
     1861 aatctccaaa gaggagaaag gtaccatgcg taaaggagaa gaacttttca ctggagttgt
     1921 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     1981 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     2041 actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc aatgctttgc
     2101 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
     2161 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     2221 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     2281 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
     2341 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
     2401 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     2461 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     2521 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     2581 catggatgaa ctatacaaat aaaagcttac gcgt
//
