LOCUS       sequence-158169-        7042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Zdk1 tagged VAV2 DH/PH/C1 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7042)
  AUTHORS   Wang H, Vilela M, Winkler A, Tarnawski M, Schlichting I, Yumerefendi
            H, Kuhlman B, Liu R, Danuser G, Hahn KM
  TITLE     LOVTRAP: an optogenetic system for photoinduced protein 
            dissociation.
  JOURNAL   Nat Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926.
  PUBMED    27427858
REFERENCE   2  (bases 1 to 7042)
  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: "Nat 
            Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926."
COMMENT     Sequence Label: pTriEx-mCherry-Zdk1-VAV2 DH/PH/C1
FEATURES             Location/Qualifiers
     source          1..7042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     110..937
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             110..234
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(272..877)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        1005..1308
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1310..1512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1463..1483
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1715..1737
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1799..1818
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1860..1879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1879..1903
                     /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        1919..2028
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             2052..2756
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2194..2212)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             2772..2948
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk1
                     /translation="MVDNKFNKEKTRAGAEIHSLPNLNVEQKFAFIVSLFDDPSQSANL
                     LAEAKKLNDAQAPK"
     CDS             4185..4217
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             4224..4247
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(4337..4356)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4402..4457
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4456..4475)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      4545..4592
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4605..5310
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           4693..5065
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(4722..5310)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(5319..5352)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(5520..6107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5599..5618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6120..6911,6912..6980))
                     /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     6743..6762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag
       61 gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta
      121 atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtcgacaag
      181 ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata
      241 aaacaattat aaatgtcaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa
      301 catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa
      361 aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac
      421 tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcttcataaa
      481 aattaacata gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt
      541 tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc
      601 tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta
      661 aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt
      721 acgcagcttc ttctagttca attacaccat tttttagcag caccggatta acataacttt
      781 ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt
      841 ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat
      901 atcacaaact ggaaatgtct atcaatatat agttgctgat ggccggccta ttaatagtaa
      961 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1021 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1081 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1141 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
     1201 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
     1261 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     1321 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1381 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1441 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1501 ataagcagac gtcgtttagt gaaccgtcag atcactagat gctttattgc ggtagtttat
     1561 cacagttaaa ttgctaacgc cagtctcgaa cttaacgtgc agaagttggt cgtgaggcac
     1621 tgggcaggta agtatcgggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     1681 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1741 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1801 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt
     1861 aatacgactc actatagggg aattgtgagc ggataacaat tccccggagt taatccggga
     1921 cctttaattc aacccaacac aatatattat agttaaataa gaattattat caaatcattt
     1981 gtatattaat taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata
     2041 ccatgtctag agtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     2101 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2161 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2221 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     2281 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     2341 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     2401 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     2461 gccctgtaat gcagaagaag actatgggct gggaggcctc ctccgagcgg atgtaccccg
     2521 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2581 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2641 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     2701 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagggtt
     2761 ctggaagttc catggtggat aacaaattca ataaagaaaa gacgcgtgcc ggtgcggaaa
     2821 tccattctct gccaaacctg aatgttgagc agaagtttgc cttcatcgtg agcctgtttg
     2881 atgatccatc tcagagcgcc aatctgctgg ccgaagccaa aaaactgaac gatgcccagg
     2941 ccccaaaagg atccggatct ggtggatctg gtaaaatggg catgactgag gacgacaaga
     3001 gaagctgctg cttgttagag attcaggaga ccgaggccaa gtactaccgc accctggagg
     3061 acattgagaa gaactacatg ggtcccttgc ggctggtgct gagcccggcg gatatggctg
     3121 ctgtcttcat caacctggag gacctcatca aggtgcatca cagctttctg cgagccatcg
     3181 atgtgtccat gatggctggt ggcagtaccc tggctaaggt ctttctggag tttaaggaaa
     3241 ggctcctgat ctatggagag tactgtagcc acatggaaca cgctcagagt acactgaacc
     3301 agctcctcgc cagccgagag gacttcaggc agaaagtgga ggagtgcaca ctcagggttc
     3361 aggatggcaa gttcaagctg caagacctgc tggtggtgcc catgcaacgg gtgctgaagt
     3421 accacctgct gctcaaggag ctcctgagcc attctgcaga ccgaccagaa agacaacagc
     3481 tcaaagaagc cctggaagcc atgcaggact tggccatgta catcaatgaa gtgaagcggg
     3541 acaaggagac cttgaagaag attagcgagt tccagtgctc catagaaaac ctgcaagtga
     3601 agctggagga atttgggagg ccaaagattg acggggagct taaagtccgg tccatagtca
     3661 accacaccaa gcaagacagg tacctgttcc tatttgacaa ggtggtcatc gtgtgcaaga
     3721 ggaagggcta cagctatgag ctgaaggagg tcattgagct gctcttccac aagatgaccg
     3781 atgacccgat gcacaacaag gacatcaaga agtggtccta tggcttctac ctgattcacc
     3841 tccaaggaaa gcaaggcttt cagttcttct gcaagacgga agacatgaag cggaagtgga
     3901 tggagcagtt cgagatggcc atgtcaaaca tcaagccaga taaggccaat gccaaccatc
     3961 atagcttcca gatgtacaca ttcgacaaga ctaccaactg caaagcctgc aagatgtttc
     4021 tcaggggtac cttctaccag ggatacctgt gtaccagatg tggcgtcggg gcacacaagg
     4081 aatgcctgga ggtgatcccc ccctgcaaga tgagttcacc tgcagatgtg gatgctcctg
     4141 gagcataaaa gcttgcggcc gcacagctgt atacacgtgc aagccagcca gaactcgccc
     4201 cggaagaccc cgaggatctc gagcaccacc atcaccatca ccatcactaa gtgattaacc
     4261 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
     4321 ataccactga gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     4381 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     4441 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     4501 atgagtattt ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg
     4561 gggcctctaa acgggtcttg aggggttttt tgctgaaagc atgcggagga aattctcctt
     4621 gaagtttccc tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt
     4681 ccggcgtatt aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg
     4741 cgttgttgat ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct
     4801 ttagggtggt atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta
     4861 aatattaaat cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt
     4921 ttttccgaac cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc
     4981 aaatcttgta gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa
     5041 ggttcgacgt cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg
     5101 tacaattgac tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg
     5161 ttagctttat taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc
     5221 gaagacgatt ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg
     5281 atcaaatttg tagttgagct ttttggaatt gcgatcgcat aacttcgtat agcatacatt
     5341 atacgaagtt ataagctcgg aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5401 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5461 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5521 ttccataggc tccgccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5701 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5941 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttgt
     6121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6421 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6481 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6541 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6601 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6661 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6721 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6781 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6841 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6901 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6961 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7021 tattgtctca tgtccgcgcg tt
//
