LOCUS       sequence-358222-        7208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression vector p5xQUAS-ZsGreen-P2A, 
            alpha-cry:mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  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 7208)
  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: p5xQUAS-ZsGreen-P2A, alpha-cry:mCherry
FEATURES             Location/Qualifiers
     source          1..7208
                     /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)"
     protein_bind    820..840
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1031..1055
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1062..1070,1071..1085)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1086..1775
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="AQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAINL
                     CVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAVC
                     ICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKGD
                     VSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEHA
                     IASGSALP"
     CDS             1791..1847
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     1853..1869
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1854..1870
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3825..3834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3831..4541
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3976..3994)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4244..4263
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4664..4785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4701..4720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4755..4774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4802..4989,4990..5001)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5020..5040)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5020..5038)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5059..5075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5059..5075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5072..5094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5083..5099
                     /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(5107..5113,5114..5131,5132..5137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5152..5173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5290..5307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5461..6049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5541..5560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6220..7011,7012..7080))
                     /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     6843..6862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7081..7185)
                     /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 aactttgtat agaaaagttg
      841 gggtaatcgc ttatcctcgg ataaacaatt atcctcacgg gtaatcgctt atccgctcgg
      901 gtaatcgctt atcctcgggt aatcgcttat ccttgcaagg gtcgactcta gagggtatat
      961 aatggatccc atcgcgtctc agcctcactt tgagctcctc cacacgaatt ccctcgacct
     1021 cgaagacgcg tcaagtttgt acaaaaaagc aggtcaccat ggatctgtac gacgatgacg
     1081 ataaggccca gtccaagcac ggcctgacca aggagatgac catgaagtac cgcatggagg
     1141 gctgcgtgga cggccacaag ttcgtgatca ccggcgaggg catcggctac cccttcaagg
     1201 gcaagcaggc catcaacctg tgcgtggtgg agggcggccc cttgcccttc gccgaggaca
     1261 tcttgtccgc cgccttcatg tacggcaacc gcgtgttcac cgagtacccc caggacatcg
     1321 tcgactactt caagaactcc tgccccgccg gctacacctg ggaccgctcc ttcctgttcg
     1381 aggacggcgc cgtgtgcatc tgcaacgccg acatcaccgt gagcgtggag gagaactgca
     1441 tgtaccacga gtccaagttc tacggcgtga acttccccgc cgacggcccc gtgatgaaga
     1501 agatgaccga caactgggag ccctcctgcg agaagatcat ccccgtgccc aagcagggca
     1561 tcttgaaggg cgacgtgagc atgtacctgc tgctgaagga cggtggccgc ttgcgctgcc
     1621 agttcgacac cgtgtacaag gccaagtccg tgccccgcaa gatgcccgac tggcacttca
     1681 tccagcacaa gctgacccgc gaggaccgca gcgacgccaa gaaccagaag tggcacctga
     1741 ccgagcacgc catcgcctcc ggctccgcct tgcccgcatg cggaagcgga gctactaact
     1801 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctaga tctcgaggtc
     1861 gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatccac tagtaattaa
     1921 gtctcagcca ccgttaactg aacatgtcaa aacctgtgga gactgttgag atttgatgtt
     1981 ctgaaaagat aaagcctata aataaaatgt tgcccaaatt tcctgcctga tgtttttctt
     2041 tgtctttgct acatggcttt gctgctcgga tcggctcact ctgtgtatgc cacgttcact
     2101 ttgtactctc cttctcacgg taggtttatt atttttagat gtgcagttag tttctgtgaa
     2161 ataacacacc acacactgat attgtctgtg cattgacttg gtgagtgcac attgtttttg
     2221 atcttgacat atttatattt gattgatcag gtgaactgtg tgaatctaaa gtgctccata
     2281 cagatgttct gcattgaaaa tattctcatt ttattagtgg aagtgagtgt atgccacatc
     2341 caatcaattt cagcaaacac cccagtatga tttaatgcaa aaaaatgaag gtatcaaaca
     2401 cgcattacta ctttgcagtt aaatatttaa catttattcc aacacgaaaa aaagcagtaa
     2461 ataacacttt gacaaacacg tcaggacatc ttatttttgt caccctcaca ggcaatttag
     2521 tataatatat tatatatata tatatatcat ataataatat tcagtataat atatatatat
     2581 atatcatatt ataatattca gtataatata aaacacaaac acatatatgt ataatataat
     2641 ataacatttt tatttattga gatgcctcta tggaccgtgt tataagaagt aaagatcagg
     2701 agaagtaaac atgaagtgta attatgaata ctgatgttaa attaagctat gatgagtttt
     2761 cactgttaat ttaccatctc aattaaatgt tgatgcctcc atgaccaagt taagcagatg
     2821 agactgagga caactgtaga agacaagatg ttcactttgc tgaatatagc tggcttgaca
     2881 gttatctatg actctataaa tatatatata tttttttttt tataaaatga tttatttata
     2941 actatatatc catttctcag acaggtgctt catatccctc actcccgtag ctgtccatgc
     3001 tggatctgtc cccgttgttt ttaaaaagct aaataagtta ttaacatgac tgcatccagc
     3061 gagccaaacc tgtctggtgt acagctacca gagaagcttg agatcctagt caccggcggc
     3121 cgctaatatg gcaataaacg atcttcagag aaacttgcat tggtaaatag tttcagccta
     3181 tgcatatgta gttattcaaa ctgtaatgac tatcttacag tattatatgt ttataatagt
     3241 ttgatccaat aaatccagat ttatatgctt ttcaacataa aattgtgaaa tgcacagaat
     3301 gtacatatac aatacatttc ctacaataat gctctctttt ctattaatag tgtgcattca
     3361 gtgcaggatg ttatgtattt attttctgta gtgcaaactt tttcaaagaa ttttccagtc
     3421 taagcttttc agtcaagaaa aaaaaagcga ttgattatct gtgatattta aaaaaattag
     3481 ctcttaaatc tggtgaacca agtggccaca acacattaaa tactgacgtt ctacagccat
     3541 gtagcttcat ctggtctggt ttgttttggc aggcacttat tttccggtga tagtcggcca
     3601 caagagtatt gtctgagcca ttcagtgcta gactgtcatt ctcaggtcag aatccacccc
     3661 actgattctg ctgacaacac tgttcccacc atgagataat gccattccag agagatccat
     3721 ttgtaagccc ctctttctgc agcacaggta tataaccagg ggtctgcctc cactaaggcc
     3781 ggcacacatc atttggggat ctttgtactg tggatccacc ggtcgccacc atggtgagca
     3841 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     3901 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3961 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     4021 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     4081 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     4141 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     4201 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     4261 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     4321 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     4381 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     4441 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     4501 gccgccactc caccggcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     4561 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4621 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4681 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4741 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaagccg cggtggagct
     4801 caagtgatct ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa
     4861 gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt
     4921 actcaagtaa agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta
     4981 ctcaagtact ttacacctct ggagctccag cttttgttcc ctttagtgag ggttaattgc
     5041 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7201 agtgccac
//
