LOCUS       sequence-358290-        6951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gene expression vector pQFDBD-2x AD*-VP16*, alpha-cry:EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6951)
  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 6951)
  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*, alpha-cry:EGFP
FEATURES             Location/Qualifiers
     source          1..6951
                     /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      3555..3564
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3561..3563,3564..3566,3567..4280)
                     /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(3561..3563,3564..3566,3567..4277)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3606..3627)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3867..3886)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4214..4235
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4407..4528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4444..4463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4498..4517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4545..4732,4733..4744)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4763..4783)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4763..4781)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4802..4818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4802..4818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4815..4837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4826..4842
                     /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(4850..4856,4857..4874,4875..4880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4895..4916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5033..5050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5204..5792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5284..5303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5963..6754,6755..6823))
                     /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     6586..6605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6824..6928)
                     /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 gctgcagtga ggggatccac tagtaattaa gtctcagcca ccgttaactg aacatgtcaa
     1681 aacctgtgga gactgttgag atttgatgtt ctgaaaagat aaagcctata aataaaatgt
     1741 tgcccaaatt tcctgcctga tgtttttctt tgtctttgct acatggcttt gctgctcgga
     1801 tcggctcact ctgtgtatgc cacgttcact ttgtactctc cttctcacgg taggtttatt
     1861 atttttagat gtgcagttag tttctgtgaa ataacacacc acacactgat attgtctgtg
     1921 cattgacttg gtgagtgcac attgtttttg atcttgacat atttatattt gattgatcag
     1981 gtgaactgtg tgaatctaaa gtgctccata cagatgttct gcattgaaaa tattctcatt
     2041 ttattagtgg aagtgagtgt atgccacatc caatcaattt cagcaaacac cccagtatga
     2101 tttaatgcaa aaaaatgaag gtatcaaaca cgcattacta ctttgcagtt aaatatttaa
     2161 catttattcc aacacgaaaa aaagcagtaa ataacacttt gacaaacacg tcaggacatc
     2221 ttatttttgt caccctcaca ggcaatttag tataatatat tatatatata tatatatcat
     2281 ataataatat tcagtataat atatatatat atatcatatt ataatattca gtataatata
     2341 aaacacaaac acatatatgt ataatataat ataacatttt tatttattga gatgcctcta
     2401 tggaccgtgt tataagaagt aaagatcagg agaagtaaac atgaagtgta attatgaata
     2461 ctgatgttaa attaagctat gatgagtttt cactgttaat ttaccatctc aattaaatgt
     2521 tgatgcctcc atgaccaagt taagcagatg agactgagga caactgtaga agacaagatg
     2581 ttcactttgc tgaatatagc tggcttgaca gttatctatg actctataaa tatatatata
     2641 tttttttttt tataaaatga tttatttata actatatatc catttctcag acaggtgctt
     2701 catatccctc actcccgtag ctgtccatgc tggatctgtc cccgttgttt ttaaaaagct
     2761 aaataagtta ttaacatgac tgcatccagc gagccaaacc tgtctggtgt acagctacca
     2821 gagaagcttg agatcctagt caccggcggc cgctaatatg gcaataaacg atcttcagag
     2881 aaacttgcat tggtaaatag tttcagccta tgcatatgta gttattcaaa ctgtaatgac
     2941 tatcttacag tattatatgt ttataatagt ttgatccaat aaatccagat ttatatgctt
     3001 ttcaacataa aattgtgaaa tgcacagaat gtacatatac aatacatttc ctacaataat
     3061 gctctctttt ctattaatag tgtgcattca gtgcaggatg ttatgtattt attttctgta
     3121 gtgcaaactt tttcaaagaa ttttccagtc taagcttttc agtcaagaaa aaaaaagcga
     3181 ttgattatct gtgatattta aaaaaattag ctcttaaatc tggtgaacca agtggccaca
     3241 acacattaaa tactgacgtt ctacagccat gtagcttcat ctggtctggt ttgttttggc
     3301 aggcacttat tttccggtga tagtcggcca caagagtatt gtctgagcca ttcagtgcta
     3361 gactgtcatt ctcaggtcag aatccacccc actgattctg ctgacaacac tgttcccacc
     3421 atgagataat gccattccag agagatccat ttgtaagccc ctctttctgc agcacaggta
     3481 tataaccagg ggtctgcctc cactaaggcc ggcacacatc atttggggat ctttgtactg
     3541 tggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3601 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3661 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3721 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3781 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3841 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3901 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3961 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4021 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4081 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4141 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4201 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4261 tggacgagct gtacaagtaa agcggccggc cgcgactcta gatcataatc agccatacca
     4321 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4381 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4441 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4501 gtttgtccaa actcatcaat gtatcttaag ccgcggtgga gctcaagtga tctccaaaaa
     4561 ataagtactt tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa
     4621 aaaatgtaat taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa
     4681 atccccaaaa ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc
     4741 tctggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     4801 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4861 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5641 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
