LOCUS       sequence-358270-        7957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression vector pQFDBD-2x AD*-VP16*-EcR, 
            alpha-cry:EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7957)
  AUTHORS   Hong J, Lee JG, Sohn KC, Lee K, Lee S, Lee J, Hong J, Choi D, Hong 
            Y, Jin HS, Choi DK, Lee SU, Kee Y, Jung J, Bae YK, Hwang RH, Hur GM,
            Lee JS, Ro H
  TITLE     IQ-Switch is a QF-based innocuous, silencing-free, and inducible 
            gene switch system in zebrafish.
  JOURNAL   Commun Biol. 2021 Dec 16;4(1):1405. doi: 10.1038/s42003-021-02923-3.
  PUBMED    34916605
REFERENCE   2  (bases 1 to 7957)
  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: "Commun 
            Biol."; date: "2021-12-16"; volume: "4(1):1405. doi"; pages: " 
            10.1038/s42003-021-02923-3"
COMMENT     Sequence Label: pQFDBD-2x AD*-VP16*-EcR, alpha-cry:EGFP
FEATURES             Location/Qualifiers
     source          1..7957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(659..670,671..808)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     810..826
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     811..827
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4561..4570
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4567..4569,4570..4572,4573..5286)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4567..4569,4570..4572,4573..5283)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4612..4633)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4873..4892)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5220..5241
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5413..5534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5450..5469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5504..5523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(5551..5738,5739..5750)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5769..5789)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5769..5787)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5808..5824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5808..5824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5821..5843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5832..5848
                     /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(join(5856..5862,5863..5880,5881..5886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5901..5922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6039..6056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6210..6798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6290..6309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6969..7760,7761..7829))
                     /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     7592..7611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7830..7934)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccca
      661 gaggtgtaaa aagtactcaa aaattttact caagtgaaag tacaagtact tagggaaaat
      721 tttactcaat taaaagtaaa agtatctggc tagaatctta cttgagtaaa agtaaaaaag
      781 tactccatta aaattgtact tgagtattct cgaggtcgac ggtatcgata agcttgatat
      841 cgaattcctg cagcccgggg ccaacatgcc gcctaaacgc aagacactca atgccgctgc
      901 cgaagccaat gcccacgctg atggccatgc tgatggcaat gctgatggtc atgtcgctaa
      961 cactgcagca agcagcaaca acgcccgttt tgcggacttg accaacattg acacacccgg
     1021 cctcggccct accacgacga cgttacttgt cgagcccgct cgttcgaaac gccagagagt
     1081 ctcgagggcc tgtgatcagt gtcgagctgc acgtgaaaag tgtgatggaa tccagccggc
     1141 ttgcttcccc tgtgtgtcgc agggccggtc gtgtacctac caggccagtc ccaagaagcg
     1201 aggagtccag acgggctaca tccgcactct cgaactggct ctggcttgga tgttcgagaa
     1261 cgttgcccgc agcgaggacg ccctccacaa tcttttggtc cgtgatgctg gccagggcag
     1321 cgctctcctg gtcggcaaag actcgcctgc tgcagaacgc ctgcatgcaa gatgggcgac
     1381 gagtcgagtc aacaaaagca tcacccgtct tctctcaggt caggccgcac aagatccatc
     1441 tgaagacggc caatccccgt ccgaagacat aaatgtccaa gatcaggccg cacaagatcc
     1501 atctgaagac ggccaatccc cgtccgaaga cataaatgtc caagatgagt tcctggatat
     1561 ggccgacttc gagtttgagc agatgtttac cgatgccctt ggaattgacg agtacggtgg
     1621 gctgcagatg aggcctgaat gtgtcataca ggagcccagt aaaaataaag acaggcaaag
     1681 acaaaagaaa gacaaaggaa tattattacc tgttagtacg accacagtcg aagaccacat
     1741 gcccccgatc atgcaatgtg atccacctcc gcccgaggcc gccaggattc acgaagtcgt
     1801 cccgaggtat ctttcggaga agctgatgga gcagaacagg cagaagaaca taccaccatt
     1861 gtcggcgaat cagaagtctc tgatcgcgag gctcgtgtgg taccaggagg gatatgagca
     1921 gccctccgac gaggatctca aaagagtaac gcagacttgg cagtcggatg aagaggacga
     1981 ggaatccgat ctacccttcc gccagatcac ggagatgacg atcttaacgg tccagttgat
     2041 cgtcgagttc gccaagggtc taccgggctt ttcgaagata tcacagtctg atcaaatcac
     2101 cttattaaaa gcctcgtcca gcgaggtgat gatgctgcgg gtggcgaggc gatacgacgc
     2161 cgcgtccgac agcattctgt tcgccaacaa caaggcgtac acgcgcgaca actaccgcaa
     2221 ggcgggcatg gccgaggtca tcgaagacct cctacacttc tgccggtgca tgttcgcgat
     2281 gggcatggac aatgtgcact ttgcactgct cacggccatc gttatattct cagatcggcc
     2341 cgggctcgag cagccgtcgc tggtagaaga gatccagaga tactacctga acacgttgcg
     2401 aatttacatc atcaaccaga acagcgcgtc gtcgcgctgc gccgtgatct acggcaggat
     2461 cctgagcgtg ctgaccgagc tacgcacgct cggcacgcaa aactccaaca tgtgcatctc
     2521 gctgaagctg aagaacagga agctgccgcc gttcctcgag gagatctggg acgtggcgga
     2581 ggtggccacg acgcatccca cggtgctgcc gcccaccaac ccggtggtgc tatagggggg
     2641 atccactagt aattaagtct cagccaccgt taactgaaca tgtcaaaacc tgtggagact
     2701 gttgagattt gatgttctga aaagataaag cctataaata aaatgttgcc caaatttcct
     2761 gcctgatgtt tttctttgtc tttgctacat ggctttgctg ctcggatcgg ctcactctgt
     2821 gtatgccacg ttcactttgt actctccttc tcacggtagg tttattattt ttagatgtgc
     2881 agttagtttc tgtgaaataa cacaccacac actgatattg tctgtgcatt gacttggtga
     2941 gtgcacattg tttttgatct tgacatattt atatttgatt gatcaggtga actgtgtgaa
     3001 tctaaagtgc tccatacaga tgttctgcat tgaaaatatt ctcattttat tagtggaagt
     3061 gagtgtatgc cacatccaat caatttcagc aaacacccca gtatgattta atgcaaaaaa
     3121 atgaaggtat caaacacgca ttactacttt gcagttaaat atttaacatt tattccaaca
     3181 cgaaaaaaag cagtaaataa cactttgaca aacacgtcag gacatcttat ttttgtcacc
     3241 ctcacaggca atttagtata atatattata tatatatata tatcatataa taatattcag
     3301 tataatatat atatatatat catattataa tattcagtat aatataaaac acaaacacat
     3361 atatgtataa tataatataa catttttatt tattgagatg cctctatgga ccgtgttata
     3421 agaagtaaag atcaggagaa gtaaacatga agtgtaatta tgaatactga tgttaaatta
     3481 agctatgatg agttttcact gttaatttac catctcaatt aaatgttgat gcctccatga
     3541 ccaagttaag cagatgagac tgaggacaac tgtagaagac aagatgttca ctttgctgaa
     3601 tatagctggc ttgacagtta tctatgactc tataaatata tatatatttt tttttttata
     3661 aaatgattta tttataacta tatatccatt tctcagacag gtgcttcata tccctcactc
     3721 ccgtagctgt ccatgctgga tctgtccccg ttgtttttaa aaagctaaat aagttattaa
     3781 catgactgca tccagcgagc caaacctgtc tggtgtacag ctaccagaga agcttgagat
     3841 cctagtcacc ggcggccgct aatatggcaa taaacgatct tcagagaaac ttgcattggt
     3901 aaatagtttc agcctatgca tatgtagtta ttcaaactgt aatgactatc ttacagtatt
     3961 atatgtttat aatagtttga tccaataaat ccagatttat atgcttttca acataaaatt
     4021 gtgaaatgca cagaatgtac atatacaata catttcctac aataatgctc tcttttctat
     4081 taatagtgtg cattcagtgc aggatgttat gtatttattt tctgtagtgc aaactttttc
     4141 aaagaatttt ccagtctaag cttttcagtc aagaaaaaaa aagcgattga ttatctgtga
     4201 tatttaaaaa aattagctct taaatctggt gaaccaagtg gccacaacac attaaatact
     4261 gacgttctac agccatgtag cttcatctgg tctggtttgt tttggcaggc acttattttc
     4321 cggtgatagt cggccacaag agtattgtct gagccattca gtgctagact gtcattctca
     4381 ggtcagaatc caccccactg attctgctga caacactgtt cccaccatga gataatgcca
     4441 ttccagagag atccatttgt aagcccctct ttctgcagca caggtatata accaggggtc
     4501 tgcctccact aaggccggca cacatcattt ggggatcttt gtactgtgga tccaccggtc
     4561 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4621 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4681 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     4741 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     4801 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4861 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4921 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4981 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     5041 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     5101 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5161 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     5221 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5281 aagtaaagcg gccggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     5341 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     5401 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5521 atcaatgtat cttaagccgc ggtggagctc aagtgatctc caaaaaataa gtactttttg
     5581 actgtaaata aaattgtaag gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag
     5641 taaaagtaaa agtattgatt tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa
     5701 tacttaagta cagtaatcaa gtaaaattac tcaagtactt tacacctctg gagctccagc
     5761 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     5821 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5881 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5941 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6001 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6061 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6121 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6181 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6241 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6301 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6361 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6421 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6481 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6541 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6601 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6661 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6721 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6781 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6841 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6901 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6961 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7021 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7081 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7141 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7201 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7261 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7321 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7381 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7441 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7501 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7561 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7621 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7681 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7741 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7801 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7861 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7921 ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
