LOCUS       Exported                8079 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes the membrane-targeted TMD-NS3-Gal4, which exhibits 
            drug-inducible transcription ""turn-off"" activity. SNAP-tag is 
            fused to the extracellular region of the encoded protein construct..
ACCESSION   .
VERSION     .
KEYWORDS    ss-SNAP-TMD-NS3-Gal4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8079)
  AUTHORS   Tague EP, Dotson HL, Tunney SN, Sloas DC, Ngo JT
  TITLE     Chemogenetic control of gene expression and cell signaling with 
            antiviral drugs.
  JOURNAL   Nat Methods. 2018 Jul;15(7):519-522. doi: 10.1038/s41592-018-0042-y.
            Epub 2018 Jul 2.
  PUBMED    29967495
REFERENCE   2  (bases 1 to 8079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8079
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1049..1594
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     CDS             1607..1636
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2636..2662
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3501..3518)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3507..3731
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3777..4205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3864..3883)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4074..4095
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4214..4234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4219..4548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4399..4534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4461..4480
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4597..5622
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5752..5873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5789..5808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5843..5862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5922..5938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5922..5938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5935..5957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5946..5962
                     /label=lac operator
                     /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)."
     promoter        complement(5970..6000)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6153..6170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6404..6423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7083..7943)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7706..7725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7944..8048)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgcggct ctgcatcccg caggtgctgt tggccttgtt
      961 cctttccatg ctgacagggc cgggagaagg cagcgctagc gatatcggcg cgccagcatt
     1021 taaatctgta cagaccggtg aattcaccat ggacaaagac tgcgaaatga agcgcaccac
     1081 cctggatagc cctctgggca agctggaact gtctgggtgc gaacagggcc tgcaccgtat
     1141 catcttcctg ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg ccccagccgc
     1201 cgtgctgggc ggaccagagc cactgatgca ggccaccgcc tggctcaacg cctactttca
     1261 ccagcctgag gccatcgagg agttccctgt gccagccctg caccacccag tgttccagca
     1321 ggagagcttt acccgccagg tgctgtggaa actgctgaaa gtggtgaagt tcggagaggt
     1381 catcagctac agccacctgg ccgccctggc cggcaatccc gccgccaccg ccgccgtgaa
     1441 aaccgccctg agcggaaatc ccgtgcccat tctgatcccc tgccaccggg tggtgcaggg
     1501 cgacctggac gtggggggct acgagggcgg gctcgccgtg aaagagtggc tgctggccca
     1561 cgagggccac agactgggca agcctgggct gggtggggga ggaagtgagc aaaagctgat
     1621 ttccgaggag gaccttggag ggggctccgc cgtggggcaa gacacccagg aggtgatcgt
     1681 ggtgcctcat agcctccctt tcaaagtggt ggtgatttca gccatcctgg cattggttgt
     1741 gctgaccatc ataagcttga ttatactgat catgctttgg cagaagaagc ccaggcgtac
     1801 gatggccagc atgactggtg gacagcaaat ggggtcgacg gaggacgtgg tgtgctgcca
     1861 ctcaatctac ggcaagaaga agggtgatat cgacacctac cgatacatag gctcttccgg
     1921 gacaggctgc gtggtcatag tgggcaggat cgtcttgtcc ggatccggca ctagtgcgcc
     1981 catcacggcg tacgcccagc agacgagagg cctcctaggg tgtataatca ccagcctgac
     2041 tggccgggac aaaaaccaag tggagggtga ggtccagatc gtgtcaactg ctacccaaac
     2101 cttcctggca acgtgcatca atggggtatg ctgggcagtc taccacgggg ccggaacgag
     2161 gaccatcgca tcacccaagg gtcctgtcat ccagatgtat accaatgtgg accaagacct
     2221 tgtgggctgg cccgctcctc aaggttcccg ctcattgaca ccctgtacct gcggctcctc
     2281 ggacctttac ctggtcacga ggcacgccga tgtcattccc gtgcgccggc gaggtgatag
     2341 caggggtagc ctgctttcgc cccggcccat ttcctacttg aaaggctcct cggggggtcc
     2401 gctgttgtgc cccgcgggac acgccgtggg cctattcagg gccgcggtgt gcacccgtgg
     2461 agtggctaaa gcggtggact ttatccctgt ggagaaccta gagacaacca tgagatcccc
     2521 ggtgttcacg gacaactcct ctccaccagc agtcaccctg acgcacccaa tcaccaaaat
     2581 cgatagggag gttctctacc aggagttcga tgagatggaa gagtgctctc agcactatcc
     2641 ctacgatgtg cccgattacg ctggcgcgtc tgcatgcggt accatgaagt tgctgagcag
     2701 catagagcaa gcatgtgata tctgccggtt gaagaagctg aagtgtagca aggagaagcc
     2761 caagtgcgcc aagtgtctca agaataattg ggagtgtagg tatagcccca agaccaagcg
     2821 aagcccgctt acgagagcac accttaccga ggtcgagagc cgcctggaaa gactcgaaca
     2881 actttttctt ctgattttcc ccagggagga cctggacatg atcctgaaga tggacagcct
     2941 ccaggacatc aaagcccttc ttaccgggct gttcgtgcag gacaacgtca acaaggatgc
     3001 ggtgaccgac agattggcga gcgtggagac ggacatgccc ttgaccctca gacaacatag
     3061 gatcagcgcg acaagctcat ctgaagaatc tagcaataag ggacagcgac agctgaccgt
     3121 tagtgccaac tttaatcaaa gtggaaacat cgcggacagc tcactcagct ttaccttcac
     3181 caatagcagt aacgggccga acctcataac cacccagacc aacagccagg ccttgagcca
     3241 gccgatcgcc tcatctaacg tgcatgataa ctttatgaac aacgagatca ccgcgagtaa
     3301 gatagacgac gggaacaaca gcaagcccct tagcccaggt tggacggacc agaccgccta
     3361 caacgctttc ggcattacga ccggcatgtt caacaccacg accatggacg atgtgtacaa
     3421 ctacctgttc gatgacgaag acacaccgcc aaaccccaaa aaagaataat ctagagggcc
     3481 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3541 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3601 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3661 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3721 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3781 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3841 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3901 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3961 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4021 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4081 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4141 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4201 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4261 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4381 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4441 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4501 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4561 gcttgtatat ccattttcgg atctgatcag cacgtgatga aaaagcctga actcaccgcg
     4621 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4681 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4741 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4801 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc
     4861 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4921 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     4981 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     5041 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     5101 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     5161 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     5221 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     5281 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     5341 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     5401 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     5461 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     5521 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     5581 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtgct acgagatttc
     5641 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5701 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5881 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7921 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7981 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8041 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
