LOCUS       sequence-132283-        3956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reporter with qacR 2 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: pES006 (pTet-qacR 2/pQacA-deGFP)
FEATURES             Location/Qualifiers
     source          1..3956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(320..337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(454..999)
                     /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(534..553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1242..1260
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1304..1315
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1896..1913
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1929..2000
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2016..2043
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2155..2176)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2504..2523
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2763..2784
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             2821..2843
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3078..3737
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3144..3163)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3758..3852
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc
       61 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
      121 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
      181 acatcagcgg tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
      241 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
      301 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
      361 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
      421 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
      481 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
      541 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
      601 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
      661 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
      721 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
      781 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
      841 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
      901 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
      961 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     1021 ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc
     1081 cccatacgat ataagttgta attctcatgt ttgacagctt atcatcgata agcttccgat
     1141 ggcgcgccga gaggctttac actttatgct tccggctggt ctcagcatcc acgcattata
     1201 acgtcgtatt agctgccttt cgtcttcaat aattcttgac atccctatca gtgatagaga
     1261 tacttcgtgc aatttttata tcaccgccag gggtacaaca agcaaagagg agaaatacta
     1321 gcatgaactt gaaagataag attttgggtg ttgctaaaga gctgtttatc aaaaacggtt
     1381 acaatgctac taccactggt gagattgtga aattgtctga atcttctaaa gggaatctat
     1441 actatcattt caaaacgaag gaaaatttgt tcttggaaat cttgaacatt caactgagta
     1501 aatatcaaga acaatggaaa aaggagcaga ttaaagctaa aactaataga gaaaagtttt
     1561 atctgtataa tgagttgtct ttgactacac aatattacta tccactacag aacgcacaga
     1621 tcgagttcta cactgaatac tataagacta actccatcaa tgaaaaacag aacaaatacc
     1681 aaaataagta tatcgatgca tatcacgtta ttttcaaaga aggtaatctt aacggtgagt
     1741 ggagtataaa tgatgttaat gccgtttcta aaattgctgc tatggctgtt ttaggtattg
     1801 ttatgttcac acatgagcaa aacatcaacg aaagaattaa gttgatgaac aagttctctc
     1861 agatcttttt gaatggtctt agtaaaggtt ctggtcacca tcatcaccac cattaatgaa
     1921 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1981 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     2041 gcgtttatag tcgggcacga ctctcaaggg catcggtcga cacagaaaag cccgcctttc
     2101 ggcgggcttt gctcgagtta ttcaccagat gcttcattag atcccggcgg cggtcacgaa
     2161 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt
     2221 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2281 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac
     2341 cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta
     2401 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2461 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2521 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2581 gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt
     2641 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2701 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2761 tacgtcgccg tccagctcga ccaggatggg aacaacgcca gtgaaaagct ccatggtata
     2821 tctccttctt aaagttaaac aaaattattg cttttaagtt ccctccaatc cttatagacc
     2881 gtgcgatcgg tctataagga ttataatcta tttttttaca tattacaaca aaaaatcttg
     2941 cgaaatttaa atgatgctat ggatgtgaga ccttgatcgg gcacgtaaga ggttccaact
     3001 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     3061 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     3121 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     3181 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     3241 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     3301 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     3361 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     3421 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     3481 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     3541 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     3601 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     3661 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     3721 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta
     3781 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     3841 attggtgaga atccaagcct cggtgagaat ccaagcctcg atcaacgtct cattttcgcc
     3901 aaaagttggc ccagggcttc ccggtatcaa cagggacacc aggatttatt tattct
//
