LOCUS       sequence-358279-        7286 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Inducible gene expression vector p9xQUAS-ZsGreen-P2A, 
            alpha-cry:mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7286)
  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 7286)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: p9xQUAS-ZsGreen-P2A, alpha-cry:mCherry
FEATURES             Location/Qualifiers
     source          1..7286
                     /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"
     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    1109..1133
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1140..1148,1149..1163)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1164..1853
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="AQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAINL
                     CVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAVC
                     ICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKGD
                     VSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEHA
                     IASGSALP"
     CDS             1869..1925
                     /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     1931..1947
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1932..1948
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3903..3912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3909..4619
                     /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(4054..4072)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4322..4341
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4742..4863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4779..4798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4833..4852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5098..5118)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5098..5116)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5137..5153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5137..5153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5150..5172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5161..5177
                     /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(5185..5191,5192..5209,5210..5215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5230..5251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5368..5385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5539..6127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5619..5638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6298..7089,7090..7158))
                     /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     6921..6940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7159..7263)
                     /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 atcctctcgg gtaatcgctt atcctcggat aaacaattat cctcacgggt
      961 aatcgcttat ccgctcgggt aatcgcttat cctcgggtaa tcgcttatcc ttgcaagggt
     1021 cgactctaga gggtatataa tggatcccat cgcgtctcag cctcactttg agctcctcca
     1081 cacgaattcc ctcgacctcg aagacgcgtc aagtttgtac aaaaaagcag gtcaccatgg
     1141 atctgtacga cgatgacgat aaggcccagt ccaagcacgg cctgaccaag gagatgacca
     1201 tgaagtaccg catggagggc tgcgtggacg gccacaagtt cgtgatcacc ggcgagggca
     1261 tcggctaccc cttcaagggc aagcaggcca tcaacctgtg cgtggtggag ggcggcccct
     1321 tgcccttcgc cgaggacatc ttgtccgccg ccttcatgta cggcaaccgc gtgttcaccg
     1381 agtaccccca ggacatcgtc gactacttca agaactcctg ccccgccggc tacacctggg
     1441 accgctcctt cctgttcgag gacggcgccg tgtgcatctg caacgccgac atcaccgtga
     1501 gcgtggagga gaactgcatg taccacgagt ccaagttcta cggcgtgaac ttccccgccg
     1561 acggccccgt gatgaagaag atgaccgaca actgggagcc ctcctgcgag aagatcatcc
     1621 ccgtgcccaa gcagggcatc ttgaagggcg acgtgagcat gtacctgctg ctgaaggacg
     1681 gtggccgctt gcgctgccag ttcgacaccg tgtacaaggc caagtccgtg ccccgcaaga
     1741 tgcccgactg gcacttcatc cagcacaagc tgacccgcga ggaccgcagc gacgccaaga
     1801 accagaagtg gcacctgacc gagcacgcca tcgcctccgg ctccgccttg cccgcatgcg
     1861 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     1921 gacctagatc tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg
     1981 ggatccacta gtaattaagt ctcagccacc gttaactgaa catgtcaaaa cctgtggaga
     2041 ctgttgagat ttgatgttct gaaaagataa agcctataaa taaaatgttg cccaaatttc
     2101 ctgcctgatg tttttctttg tctttgctac atggctttgc tgctcggatc ggctcactct
     2161 gtgtatgcca cgttcacttt gtactctcct tctcacggta ggtttattat ttttagatgt
     2221 gcagttagtt tctgtgaaat aacacaccac acactgatat tgtctgtgca ttgacttggt
     2281 gagtgcacat tgtttttgat cttgacatat ttatatttga ttgatcaggt gaactgtgtg
     2341 aatctaaagt gctccataca gatgttctgc attgaaaata ttctcatttt attagtggaa
     2401 gtgagtgtat gccacatcca atcaatttca gcaaacaccc cagtatgatt taatgcaaaa
     2461 aaatgaaggt atcaaacacg cattactact ttgcagttaa atatttaaca tttattccaa
     2521 cacgaaaaaa agcagtaaat aacactttga caaacacgtc aggacatctt atttttgtca
     2581 ccctcacagg caatttagta taatatatta tatatatata tatatcatat aataatattc
     2641 agtataatat atatatatat atcatattat aatattcagt ataatataaa acacaaacac
     2701 atatatgtat aatataatat aacattttta tttattgaga tgcctctatg gaccgtgtta
     2761 taagaagtaa agatcaggag aagtaaacat gaagtgtaat tatgaatact gatgttaaat
     2821 taagctatga tgagttttca ctgttaattt accatctcaa ttaaatgttg atgcctccat
     2881 gaccaagtta agcagatgag actgaggaca actgtagaag acaagatgtt cactttgctg
     2941 aatatagctg gcttgacagt tatctatgac tctataaata tatatatatt ttttttttta
     3001 taaaatgatt tatttataac tatatatcca tttctcagac aggtgcttca tatccctcac
     3061 tcccgtagct gtccatgctg gatctgtccc cgttgttttt aaaaagctaa ataagttatt
     3121 aacatgactg catccagcga gccaaacctg tctggtgtac agctaccaga gaagcttgag
     3181 atcctagtca ccggcggccg ctaatatggc aataaacgat cttcagagaa acttgcattg
     3241 gtaaatagtt tcagcctatg catatgtagt tattcaaact gtaatgacta tcttacagta
     3301 ttatatgttt ataatagttt gatccaataa atccagattt atatgctttt caacataaaa
     3361 ttgtgaaatg cacagaatgt acatatacaa tacatttcct acaataatgc tctcttttct
     3421 attaatagtg tgcattcagt gcaggatgtt atgtatttat tttctgtagt gcaaactttt
     3481 tcaaagaatt ttccagtcta agcttttcag tcaagaaaaa aaaagcgatt gattatctgt
     3541 gatatttaaa aaaattagct cttaaatctg gtgaaccaag tggccacaac acattaaata
     3601 ctgacgttct acagccatgt agcttcatct ggtctggttt gttttggcag gcacttattt
     3661 tccggtgata gtcggccaca agagtattgt ctgagccatt cagtgctaga ctgtcattct
     3721 caggtcagaa tccaccccac tgattctgct gacaacactg ttcccaccat gagataatgc
     3781 cattccagag agatccattt gtaagcccct ctttctgcag cacaggtata taaccagggg
     3841 tctgcctcca ctaaggccgg cacacatcat ttggggatct ttgtactgtg gatccaccgg
     3901 tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     3961 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     4021 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     4081 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     4141 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     4201 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     4261 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     4321 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     4381 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     4441 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4501 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4561 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaaa
     4621 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4681 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4741 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4801 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4861 ttaagccgcg gtggagctca agtgatctcc aaaaaataag tactttttga ctgtaaataa
     4921 aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa
     4981 gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac
     5041 agtaatcaag taaaattact caagtacttt acacctctgg agctccagct tttgttccct
     5101 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5161 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5221 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5281 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5941 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7261 gcgcacattt ccccgaaaag tgccac
//
