LOCUS       sequence-310842-        7843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tol2 vector containing QUASM1 reporter element upstream of 
            mScarlet-NLS. Includes cmlc2:mRFP-SV40pA transgenesis marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7843)
  AUTHORS   Burgess J, Burrows JT, Sadhak R, Chiang S, Weiss A, D'Amata C, 
            Molinaro AM, Zhu S, Long M, Hu C, Krause HM, Pearson BJ
  TITLE     An optimized QF-binary expression system for use in zebrafish.
  JOURNAL   Dev Biol. 2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007.
  PUBMED    32697972
REFERENCE   2  (bases 1 to 7843)
  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: "Dev Biol. 
            2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007."
COMMENT     Sequence Label: pTol2CG2-QUASM1:mScarletNLS-SV40pA
FEATURES             Location/Qualifiers
     source          1..7843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(33..57)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    59..166
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     76..92
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(126..142)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(180..200)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(208..224)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    240..374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1472..1481
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1526..1535
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1589..1598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1595..2272
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1595..2272
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     1993..2012
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2187..2210
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    join(2735..2922,2923..2934)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2968..2984)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3021..3041)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3021..3039)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3060..3076)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3073..3095)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3084..3100
                     /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(3108..3114,3115..3132,3133..3138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3153..3174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3291..3308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3462..4050)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3542..3561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4221..5012,5013..5081))
                     /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     4844..4863
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5082..5186)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5349..5370)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5823..5834,5835..5997)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     6370..6386
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6406..6426
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    6436..6543
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     6453..6469
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6503..6519)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6550..6574
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      6832..6841
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(join(7833..7843,1..6))
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acctcgaggg ggggcccggt acccagcttt tgacccagct ttcttgtaca aagtgggagg
       61 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc
      121 cgggggatcc actagttcta gagcggccgc caccgcggtg gagctcaccg gtagatctgc
      181 aactttatta tacatagttg ataattcact ggccgtcgtt ttacggtacc atcgatgatg
      241 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      301 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      361 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
      421 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc
      481 ctctagatca gatcttgacc aaagcttaaa tcagttgtgt taaataagag acattcaaaa
      541 taaatgtaaa tgagctctcc aaatcagcag acttaacatt ctttaaaatg attgattcaa
      601 tagtgataaa aatcaggcat agccagttgt aactttagat aaattacaga aaatgtcaaa
      661 tacagagaac cgattctttt ttatgataca tccaagcaca catttaacac aatccaggca
      721 aaccccgaat ttcacagtca caagcactgt ttgtacaaga gctttgccta aggacacaca
      781 gtctctataa gtccaggtcg ttggtttcac tcttatttta aacatgtgac atttttcctg
      841 ccatcctgtc ttaggctgct gtttgcttca ttccatgtca cattaatttc ctcagtagca
      901 ccttttacac acacagccaa tcttttccag aaaattcaat tgctttgaag agataatgtg
      961 tgaacaaatc catttagaaa aggaaaatta agaatttgta aaatcatctg taaattgttg
     1021 gcattcttct gtatatgaac atcacatcat ttacaggtaa aggtctggtc attaattata
     1081 tgacaattta ctggtattat tttgtgaaag gggctatttt caatgcgttc atccatcctt
     1141 ttcatccctc aaatctctca ttcacgtccc cctccccatc tgcacacttt atctcatttt
     1201 ccaccctgct ggaatctgag cacttgtgca gttatcaggg ctcctgtatt taggaggctc
     1261 tgggtgtcca tgtaggggac gaacagaaac actgcagacc tttatagaag aacaaatgat
     1321 aagagtcctc gtacataaag actccattag taacgtcagt gacccaggag cccagaccaa
     1381 cagcaaagca gacagtgaaa tcactagtga attcgcggcc gcctgcaggt cgacggtatc
     1441 gattcgaatt caaggcctct cgagcctcta agccaccatg ggctgcatca agagcaagcg
     1501 caaggacaac ctgaacgacg acgaggccgc catgggctgc atcaagagca agcgcaagga
     1561 caacctgaac gacgacgagg caccggtcgc caccatggcc tcctccgagg acgtcatcaa
     1621 ggagttcatg cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat
     1681 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1741 caagggcggc cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc
     1801 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     1861 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     1921 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     1981 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     2041 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga
     2101 cggcggccac tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct
     2161 gcccggcgcc tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac
     2221 catcgtggaa cagtacgagc gcgccgaggg ccgccactcc accggcgcct aatacgtaga
     2281 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2341 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2401 caataaacaa gagatctgcg aagatacggc cacgggtgct cttgatcctg tggctgattt
     2461 tggactgtgc tgctcgcagc tgctgatgaa tcacatactt cctccatttt cttccactga
     2521 ttgactgtta taatttccct aatttccagg tcaaggtgct gtgcattgtg gtaatagatg
     2581 tgacatgacg tcacttccaa aggaccaatg aacatgtctg accaatttca tataatgtga
     2641 aaacgatttt cataggcaga ataaataaca tttaaattaa actgggcatc agcgcaattc
     2701 aattggtttg gtaatagcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt
     2761 tttgactgta aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat
     2821 taagtaaaag taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa
     2881 ataatactta agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt
     2941 gaccccctac cttcagcaag cccagcagat ccactagttc tagagcggcc gccaccgcgg
     3001 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3061 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3121 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3181 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3241 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3301 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3361 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3421 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3481 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3541 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3601 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3661 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3721 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3781 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3841 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3901 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3961 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4021 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4081 gacgctcagt ggaacgaaaa ctcacgttaa ggcattttgg tcatgagatt atcaaaaagg
     4141 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4201 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4261 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4321 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4381 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4441 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4501 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4561 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4621 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4681 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4741 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4801 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4861 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4921 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4981 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5041 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5101 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5161 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     5221 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5281 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5341 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5401 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5461 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5521 gagcttgacg gggaaaggct ggctaagaac tcatcagcct ccccggtcca tctacccacg
     5581 taccaatgca ccaattggcc acaatgacgg ctactacatg gtgccattcc ttcctcttta
     5641 taggaatgga gactacctcc tgtccaacaa tgctcttgga tacgagtacg cctacctgtt
     5701 ggacccaggt cattgcacaa caccagaaat gccctctgat ctgcaaaaga cgtgaatatc
     5761 tgttcagaca cccatatcca ctctgttcca cacaggtcag aggtttgtcc aggagttctt
     5821 gacagaggtg taaaaagtac tcaaaaattt tactcaagtg aaagtacaag tacttaggga
     5881 aaattttact caattaaaag taaaagtatc tggctagaat cttacttgag taaaagtaaa
     5941 aaagtactcc attaaaattg tacttgagta ttaaggaagt aaaagtaaaa gcaagaaaga
     6001 aaactagaga ttcttgttta agcttttaat ctcaaaaaac attaaatgaa atgcatacaa
     6061 ggttttatcc tgctttagaa ctgtttgtat ttaattatca aactataaga cagacaatct
     6121 aatgccagta cacgctactc aaagttgtaa aacctcagat ttaacttcag tagaagctga
     6181 ttctcaaaat tgttagtgtc aagcctagct cttttggggc tgaaaagcaa tcctgcagtg
     6241 ctgaaaagcc tctcacaggc agccgatgcg ggaagaggtg tattagtctt gatagagagg
     6301 ctgcaaatag caggaaacgt gagcagagac tccctggtgt ctgaaacaca ggccagatgg
     6361 gccctcgagc aggaaacagc tatgaccatg attacgccaa gctatcaact ttgtatagaa
     6421 aagttgcaaa agctgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat
     6481 atcgaattcc tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag
     6541 ctccaattcg caagtttgta caaaaaagca ggctcgaatt ggagctccac cgcggtggcg
     6601 gccgcaagga taaacaatta tcctcacggg taatcgctta tcctcaagga taaacaatta
     6661 tcctcacggg taatcgctta tcctcaagga taaacaatta tcctcgcaag ggtcgactct
     6721 agagggtata taatggatcc catcgcgtct cagcctcact ttgagctcct ccacacgaat
     6781 tccctcgacc tcgaagacgc gtctgtttgg gatccaccgg tcactagtgc cgccaccatg
     6841 gtgagcaagg gcgaggccgt gatcaaagaa ttcatgcgct tcaaggtgca catggaaggc
     6901 agcatgaacg gacacgagtt cgagatcgaa ggcgaaggcg agggcagacc ttacgaggga
     6961 acacagaccg ccaagctgaa agtgaccaaa ggcggacctc tgcctttcag ctgggacatc
     7021 ctgtctccac agtttatgta cggaagccgc gccttcacaa agcaccctgc tgacatccct
     7081 gactactaca agcagagctt ccctgaggga ttcaagtggg agagagtgat gaacttcgag
     7141 gacggcggag ccgtgacagt gacacaggat accagcctgg aagatggaac cctgatctac
     7201 aaagtgaagc tgcgcggaac aaacttccct cctgacggac ctgtgatgca gaaaaagacc
     7261 atgggatggg aagccagcac cgagagactg taccctgaag atggcgtgct gaagggcgac
     7321 atcaagatgg ccctgagact gaaagacggc ggaagatacc tggccgactt caagaccacc
     7381 tacaaggcca agaaacctgt gcagatgcct ggcgcctaca acgtggacag aaagctggac
     7441 atcaccagcc acaacgagga ctacaccgtg gtggaacagt acgagagaag cgagggacgc
     7501 cacagcacag gcggaatgga cgagctgtac aagagcaggc acagaaggca tcgccagcgc
     7561 tctaggagcc gcaatcgcag ccgaagtcgc agcagtgaac gaaaacgccg tcaacggagc
     7621 cgaagtcgca gcagtgaacg aagacgctac ttgtacaagt aatctagaac tatagtgagt
     7681 cgtattacgt agatccagac atgataagat acattgatga gtttggacaa accacaacta
     7741 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     7801 ccattataag ctgcaataaa caagaagctt atcgataccg tcg
//
