LOCUS       Exported                5701 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  QacR-Q1 gate.
ACCESSION   .
VERSION     .
KEYWORDS    pQacR-Q1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5701)
  AUTHORS   Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski
            EA, Ross D, Densmore D, Voigt CA
  TITLE     Genetic circuit design automation.
  JOURNAL   Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341.
  PUBMED    27034378
REFERENCE   2  (bases 1 to 5701)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5701
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(308..852)
                     /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(388..407)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(941..1756)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1664..1683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1748..1767
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        1878..1906
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1912..1931
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        2752..2801
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             2851..3570
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2896..2917)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3157..3176)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3504..3525
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3718..4800
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3737..3756)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4819..5442
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4892..4911)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(5625..5644)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcttcctcgc tcactgactc gctgcacgag gcagacctca gcgctagcgg agtgtatact
       61 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      121 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      181 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      241 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      301 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      361 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct
      421 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg
      481 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact
      541 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg
      601 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta
      661 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt
      721 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc
      781 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca
      841 aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaaatca
      901 atctaaagta tatatgagta aacttggtct gacagttacc ttagaaaaac tcatcgagca
      961 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1021 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     1081 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     1141 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     1201 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1261 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1321 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1381 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1441 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1501 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1561 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1621 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1681 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     1741 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     1801 ttcatgatga tatattttta tcttgtgcaa tgtacatcag agattttgag acacaaggag
     1861 aacgatcgtt ggctgtgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     1921 cgctcacaat tagtcgtcaa gtgctgtgct tgcacttctg atgaggcagt gatgccgaaa
     1981 cgacctctac aaataatttt gtttaagtaa gccatgccat tggcttttga tagaggataa
     2041 ctactagatg aacctgaaag ataaaattct gggcgttgcc aaagaactgt ttatcaaaaa
     2101 tggctataac gcaaccacca ccggtgaaat tgttaaactg agcgaaagca gcaaaggcaa
     2161 tctgtattat cactttaaaa ccaaagagaa cctgtttctg gaaatcctga acatcgaaga
     2221 aagcaaatgg caagagcagt ggaaaaaaga acaaatcaaa tgcaaaacca accgcgagaa
     2281 attctatctg tataatgaac tgagcctgac caccgaatat tactatccgc tgcagaatgc
     2341 catcatcgag ttttataccg agtactataa aaccaacagc atcaacgaga aaatgaacaa
     2401 actggaaaac aaatacatcg atgcctacca cgtgatcttt aaagaaggta atctgaacgg
     2461 cgaatggtgc attaatgatg ttaatgccgt gagcaaaatt gcagcaaatg ccgttaatgg
     2521 cattgttacc tttacccatg agcagaatat caacgaacgc attaaactga tgaacaaatt
     2581 cagccagatc tttctgaatg gcctgagcaa ataagtcagt ttcacctgtt ttacgtaaaa
     2641 acccgcttcg gcgggttttt acttttggta ctggtatgga agctatacgt taccaattga
     2701 cagctagctc agtcctactt tagtatatag accgtgcgat cggtctataa gctgtcaccg
     2761 gatgtgcttt ccggtctgat gagtccgtga ggacgaaaca gcctctacaa ataattttgt
     2821 ttaatactag agaaagaggg gaaatactag atggtgagca agggcgagga gctgttcacc
     2881 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     2941 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3001 acaggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgcaa
     3061 tgcttcgccc gctaccccga ccacatgaag ctgcacgact tcttcaagtc cgccatgccc
     3121 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3181 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3241 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     3301 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3361 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac cccaatcggc
     3421 gacggccccg tgctgctgcc cgacaaccac taccttagct accagtccgc cctgagcaaa
     3481 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3541 actctcggca tggacgagct gtacaagtaa ctcggtacca aattccagaa aagaggcctc
     3601 ccgaaagggg ggcctttttt cgttttggtc caatggcggc gcgccatcga atggcgcaaa
     3661 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatatg
     3721 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     3781 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     3841 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     3901 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     3961 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     4021 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     4081 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     4141 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     4201 tcccatgagg acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     4261 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     4321 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     4381 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     4441 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     4501 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga tagctcatgt
     4561 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     4621 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgccagtc
     4681 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     4741 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4801 taatccagga ggaaaaaaat gtccagatta gataaaagta aagtgattaa cagcgcatta
     4861 gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta
     4921 ggtgtagagc agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc
     4981 ttagccattg agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc
     5041 tggcaagatt ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc
     5101 gatggagcaa aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa
     5161 aatcaattag cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc
     5221 agcgctgtgg ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct
     5281 aaagaagaaa gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc
     5341 gaattatttg atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata
     5401 tgcggattag aaaaacaact taaatgtgaa agtgggtcct aataattggt aacgaatcag
     5461 acaattgacg gctcgaggga gtagcatagg gtttgcagaa tccctgcttc gtccatttga
     5521 caggcacatt atgcatcgat gataagctgt caaacatgag cagatcctct acgccggacg
     5581 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     5641 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcaaatatt ttatctgagg
     5701 t
//
