LOCUS       sequence-121996-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter with wild-type qacR on single plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3956)
  AUTHORS   de Los Santos EL, Meyerowitz JT, Mayo SL, Murray RM
  TITLE     Engineering Transcriptional Regulator Effector Specificity Using 
            Computational Design and In Vitro Rapid Prototyping: Developing a 
            Vanillin Sensor.
  JOURNAL   ACS Synth Biol. 2015 Aug 19.
  PUBMED    26262913
REFERENCE   2  (bases 1 to 3956)
  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: "ACS Synth 
            Biol. 2015 Aug 19."
COMMENT     Sequence Label: pES005 (pTet-qacR wt/pQacA-deGFP)
FEATURES             Location/Qualifiers
     source          1..3956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             121..780
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(187..206)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      801..895
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1319..1336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1453..1998)
                     /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(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2241..2259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2303..2314
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2895..2912
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2928..2999
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3015..3042
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3154..3175)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3503..3522
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3762..3783
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             3820..3842
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tatgaagctg agaccttgat cgggcacgta agaggttcca actttcacca taatgaaata
       61 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
      121 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
      181 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
      241 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
      301 cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt
      361 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
      421 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
      481 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
      541 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
      601 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
      661 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
      721 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
      781 tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
      841 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
      901 cctcggtgag aatccaagcc tcgatcaacg tctcattttc gccaaaagtt ggcccagggc
      961 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     1021 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     1081 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     1141 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcggt agcggagtgt
     1201 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     1261 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     1321 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     1381 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     1441 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     1501 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     1561 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     1621 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     1681 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     1741 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     1801 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     1861 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     1921 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     1981 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca
     2041 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     2101 ttctcatgtt tgacagctta tcatcgataa gcttccgatg gcgcgccgag aggctttaca
     2161 ctttatgctt ccggctggtc tcagcatcca cgcattataa cgtcgtatta gctgcctttc
     2221 gtcttcaata attcttgaca tccctatcag tgatagagat acttcgtgca atttttatat
     2281 caccgccagg ggtacaacaa gcaaagagga gaaatactag catgaacttg aaagataaga
     2341 ttttgggtgt tgctaaagag ctgtttatca aaaacggtta caatgctact accactggtg
     2401 agattgtgaa attgtctgaa tcttctaaag ggaatctata ctatcatttc aaaacgaagg
     2461 aaaatttgtt cttggaaatc ttgaacattg aagagagtaa atggcaagaa caatggaaaa
     2521 aggagcagat taaagctaaa actaatagag aaaagtttta tctgtataat gagttgtctt
     2581 tgactacaga gtattactat ccactacaga acgcaataat cgagttctac actgaatact
     2641 ataagactaa ctccatcaat gaaaaaatga acaaattaga gaataagtat atcgatgcat
     2701 atcacgttat tttcaaagaa ggtaatctta acggtgagtg gagtataaat gatgttaatg
     2761 ccgtttctaa aattgctgct aacgctgtta acggtattgt tacattcaca catgagcaaa
     2821 acatcaacga aagaattaag ttgatgaaca agttctctca gatctttttg aatggtctta
     2881 gtaaaggttc tggtcaccat catcaccacc attgataaac caggcatcaa ataaaacgaa
     2941 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     3001 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagt cgggcacgac
     3061 tctcaagggc atcggtcgac acagaaaagc ccgcctttcg gcgggctttg ctcgagttat
     3121 tcaccagatg cttcattaga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
     3181 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg
     3241 cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac
     3301 gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt
     3361 gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat
     3421 gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc
     3481 ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg
     3541 ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc
     3601 ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg
     3661 cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc
     3721 gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac
     3781 caggatggga acaacgccag tgaaaagctc catggtatat ctccttctta aagttaaaca
     3841 aaattattgc ttttaagttc cctccaatcc ttatagaccg tgcgatcggt ctataaggat
     3901 tataatctat ttttttacat attacaacaa aaaatcttgc gaaatttaaa tgatgc
//
