LOCUS       sequence-358218-        7360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible gene expression vector p13xQUAS-ZsGreen-P2A, 
            alpha-cry:mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7360)
  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 7360)
  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: p13xQUAS-ZsGreen-P2A, alpha-cry:mCherry
FEATURES             Location/Qualifiers
     source          1..7360
                     /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    1183..1207
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1214..1222,1223..1237)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1238..1927
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="AQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAINL
                     CVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAVC
                     ICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKGD
                     VSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEHA
                     IASGSALP"
     CDS             1943..1999
                     /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     2005..2021
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2006..2022
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3977..3986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3983..4693
                     /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(4128..4146)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4396..4415
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4816..4937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4853..4872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4907..4926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(4954..5141,5142..5153)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5172..5192)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5172..5190)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5211..5227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5211..5227)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5224..5246)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5235..5251
                     /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(5259..5265,5266..5283,5284..5289))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5304..5325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5442..5459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5613..6201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5693..5712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6372..7163,7164..7232))
                     /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     6995..7014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7233..7337)
                     /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 cctcgggtaa tcgcttatcc tcggataaac
      961 aattatcctc acgggtaatc gcttatccgc tcgggtaatc gcttatcctc ggataaacaa
     1021 ttatcctcac gggtaatcgc ttatccgctc gggtaatcgc ttatcctcgg gtaatcgctt
     1081 atccttgcaa gggtcgactc tagagggtat ataatggatc ccatcgcgtc tcagcctcac
     1141 tttgagctcc tccacacgaa ttccctcgac ctcgaagacg cgtcaagttt gtacaaaaaa
     1201 gcaggtcacc atggatctgt acgacgatga cgataaggcc cagtccaagc acggcctgac
     1261 caaggagatg accatgaagt accgcatgga gggctgcgtg gacggccaca agttcgtgat
     1321 caccggcgag ggcatcggct accccttcaa gggcaagcag gccatcaacc tgtgcgtggt
     1381 ggagggcggc cccttgccct tcgccgagga catcttgtcc gccgccttca tgtacggcaa
     1441 ccgcgtgttc accgagtacc cccaggacat cgtcgactac ttcaagaact cctgccccgc
     1501 cggctacacc tgggaccgct ccttcctgtt cgaggacggc gccgtgtgca tctgcaacgc
     1561 cgacatcacc gtgagcgtgg aggagaactg catgtaccac gagtccaagt tctacggcgt
     1621 gaacttcccc gccgacggcc ccgtgatgaa gaagatgacc gacaactggg agccctcctg
     1681 cgagaagatc atccccgtgc ccaagcaggg catcttgaag ggcgacgtga gcatgtacct
     1741 gctgctgaag gacggtggcc gcttgcgctg ccagttcgac accgtgtaca aggccaagtc
     1801 cgtgccccgc aagatgcccg actggcactt catccagcac aagctgaccc gcgaggaccg
     1861 cagcgacgcc aagaaccaga agtggcacct gaccgagcac gccatcgcct ccggctccgc
     1921 cttgcccgca tgcggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt
     1981 ggaggagaac cctggaccta gatctcgagg tcgacggtat cgataagctt gatatcgaat
     2041 tcctgcagcc cgggggatcc actagtaatt aagtctcagc caccgttaac tgaacatgtc
     2101 aaaacctgtg gagactgttg agatttgatg ttctgaaaag ataaagccta taaataaaat
     2161 gttgcccaaa tttcctgcct gatgtttttc tttgtctttg ctacatggct ttgctgctcg
     2221 gatcggctca ctctgtgtat gccacgttca ctttgtactc tccttctcac ggtaggttta
     2281 ttatttttag atgtgcagtt agtttctgtg aaataacaca ccacacactg atattgtctg
     2341 tgcattgact tggtgagtgc acattgtttt tgatcttgac atatttatat ttgattgatc
     2401 aggtgaactg tgtgaatcta aagtgctcca tacagatgtt ctgcattgaa aatattctca
     2461 ttttattagt ggaagtgagt gtatgccaca tccaatcaat ttcagcaaac accccagtat
     2521 gatttaatgc aaaaaaatga aggtatcaaa cacgcattac tactttgcag ttaaatattt
     2581 aacatttatt ccaacacgaa aaaaagcagt aaataacact ttgacaaaca cgtcaggaca
     2641 tcttattttt gtcaccctca caggcaattt agtataatat attatatata tatatatatc
     2701 atataataat attcagtata atatatatat atatatcata ttataatatt cagtataata
     2761 taaaacacaa acacatatat gtataatata atataacatt tttatttatt gagatgcctc
     2821 tatggaccgt gttataagaa gtaaagatca ggagaagtaa acatgaagtg taattatgaa
     2881 tactgatgtt aaattaagct atgatgagtt ttcactgtta atttaccatc tcaattaaat
     2941 gttgatgcct ccatgaccaa gttaagcaga tgagactgag gacaactgta gaagacaaga
     3001 tgttcacttt gctgaatata gctggcttga cagttatcta tgactctata aatatatata
     3061 tatttttttt tttataaaat gatttattta taactatata tccatttctc agacaggtgc
     3121 ttcatatccc tcactcccgt agctgtccat gctggatctg tccccgttgt ttttaaaaag
     3181 ctaaataagt tattaacatg actgcatcca gcgagccaaa cctgtctggt gtacagctac
     3241 cagagaagct tgagatccta gtcaccggcg gccgctaata tggcaataaa cgatcttcag
     3301 agaaacttgc attggtaaat agtttcagcc tatgcatatg tagttattca aactgtaatg
     3361 actatcttac agtattatat gtttataata gtttgatcca ataaatccag atttatatgc
     3421 ttttcaacat aaaattgtga aatgcacaga atgtacatat acaatacatt tcctacaata
     3481 atgctctctt ttctattaat agtgtgcatt cagtgcagga tgttatgtat ttattttctg
     3541 tagtgcaaac tttttcaaag aattttccag tctaagcttt tcagtcaaga aaaaaaaagc
     3601 gattgattat ctgtgatatt taaaaaaatt agctcttaaa tctggtgaac caagtggcca
     3661 caacacatta aatactgacg ttctacagcc atgtagcttc atctggtctg gtttgttttg
     3721 gcaggcactt attttccggt gatagtcggc cacaagagta ttgtctgagc cattcagtgc
     3781 tagactgtca ttctcaggtc agaatccacc ccactgattc tgctgacaac actgttccca
     3841 ccatgagata atgccattcc agagagatcc atttgtaagc ccctctttct gcagcacagg
     3901 tatataacca ggggtctgcc tccactaagg ccggcacaca tcatttgggg atctttgtac
     3961 tgtggatcca ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat
     4021 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     4081 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4141 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     4201 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     4261 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     4321 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     4381 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     4441 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     4501 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     4561 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     4621 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     4681 gctgtacaag taaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     4741 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4801 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4921 ctcatcaatg tatcttaagc cgcggtggag ctcaagtgat ctccaaaaaa taagtacttt
     4981 ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt
     5041 aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa
     5101 taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct ctggagctcc
     5161 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     5221 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5281 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5341 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5401 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5461 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5521 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5581 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5641 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5701 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5761 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5821 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5881 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5941 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6001 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6061 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6121 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7321 caaatagggg ttccgcgcac atttccccga aaagtgccac
//
