LOCUS       sequence-154112-        7048 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 7048)
  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 7048)
  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-mVenus-Zdk1-VAV2 DH/PH/C1
FEATURES             Location/Qualifiers
     source          1..7048
                     /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             join(2043..2045,2046..2048,2049..2759)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2088..2109)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2349..2368)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2696..2717
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2778..2954
                     /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             4191..4223
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             4230..4253
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(4343..4362)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4408..4463
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4462..4481)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      4551..4598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     4611..5316
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           4699..5071
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(4728..5316)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(5325..5358)
                     /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(5526..6113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5605..5624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6126..6917,6918..6986))
                     /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     6749..6768
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2101 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2161 acggcaagct gaccctgaag ttcatcagca ccaccggcaa gctgcccgtg ccctggccca
     2221 ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     2281 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2341 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2401 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2461 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     2521 acggcatcaa ggcaaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     2581 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2641 actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc gatcacatgg
     2701 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     2761 gttctggaag tggatctatg gtggataaca aattcaataa agaaaagacg cgtgccggtg
     2821 cggaaatcca ttctctgcca aacctgaatg ttgagcagaa gtttgccttc atcgtgagcc
     2881 tgtttgatga tccatctcag agcgccaatc tgctggccga agccaaaaaa ctgaacgatg
     2941 cccaggcccc aaaaggatcc ggatctggtg gatctggtaa aatgggcatg actgaggacg
     3001 acaagagaag ctgctgcttg ttagagattc aggagaccga ggccaagtac taccgcaccc
     3061 tggaggacat tgagaagaac tacatgggtc ccttgcggct ggtgctgagc ccggcggata
     3121 tggctgctgt cttcatcaac ctggaggacc tcatcaaggt gcatcacagc tttctgcgag
     3181 ccatcgatgt gtccatgatg gctggtggca gtaccctggc taaggtcttt ctggagttta
     3241 aggaaaggct cctgatctat ggagagtact gtagccacat ggaacacgct cagagtacac
     3301 tgaaccagct cctcgccagc cgagaggact tcaggcagaa agtggaggag tgcacactca
     3361 gggttcagga tggcaagttc aagctgcaag acctgctggt ggtgcccatg caacgggtgc
     3421 tgaagtacca cctgctgctc aaggagctcc tgagccattc tgcagaccga ccagaaagac
     3481 aacagctcaa agaagccctg gaagccatgc aggacttggc catgtacatc aatgaagtga
     3541 agcgggacaa ggagaccttg aagaagatta gcgagttcca gtgctccata gaaaacctgc
     3601 aagtgaagct ggaggaattt gggaggccaa agattgacgg ggagcttaaa gtccggtcca
     3661 tagtcaacca caccaagcaa gacaggtacc tgttcctatt tgacaaggtg gtcatcgtgt
     3721 gcaagaggaa gggctacagc tatgagctga aggaggtcat tgagctgctc ttccacaaga
     3781 tgaccgatga cccgatgcac aacaaggaca tcaagaagtg gtcctatggc ttctacctga
     3841 ttcacctcca aggaaagcaa ggctttcagt tcttctgcaa gacggaagac atgaagcgga
     3901 agtggatgga gcagttcgag atggccatgt caaacatcaa gccagataag gccaatgcca
     3961 accatcatag cttccagatg tacacattcg acaagactac caactgcaaa gcctgcaaga
     4021 tgtttctcag gggtaccttc taccagggat acctgtgtac cagatgtggc gtcggggcac
     4081 acaaggaatg cctggaggtg atccccccct gcaagatgag ttcacctgca gatgtggatg
     4141 ctcctggagc ataaaagctt gcggccgcac agctgtatac acgtgcaagc cagccagaac
     4201 tcgccccgga agaccccgag gatctcgagc accaccatca ccatcaccat cactaagtga
     4261 ttaacctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     4321 tcacaaatac cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga
     4381 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     4441 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     4501 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgtaa ctagcataac
     4561 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaagcatgc ggaggaaatt
     4621 ctccttgaag tttccctggt gttcaaagta aaggagtttg caccagacgc acctctgttc
     4681 actggtccgg cgtattaaaa cacgatacat tgttattagt acatttatta agcgctagat
     4741 tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc attaaattta
     4801 taatctttag ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc tttatatctg
     4861 aatttaaata ttaaatcctc aatagatttg taaaataggt ttcgattagt ttcaaacaag
     4921 ggttgttttt ccgaaccgat ggctggacta tctaatggat tttcgctcaa cgccacaaaa
     4981 cttgccaaat cttgtagcag caatctagct ttgtcgatat tcgtttgtgt tttgttttgt
     5041 aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg tatttctttc atcactgtcg
     5101 ttagtgtaca attgactcga cgtaaacacg ttaaatagag cttggacata tttaacatcg
     5161 ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt
     5221 gcttccgaag acgattttgc catagccaca cgacgcctat taattgtgtc ggctaacacg
     5281 tccgcgatca aatttgtagt tgagcttttt ggaattgcga tcgcataact tcgtatagca
     5341 tacattatac gaagttataa gctcggaacg ctgcgctcgg tcgttcggct gcggcgagcg
     5401 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5521 gcgtttttcc ataggctccg ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5701 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     5761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5941 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6121 ctttgttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6181 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6241 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6301 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6361 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6421 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6481 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6541 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6601 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6661 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6721 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6781 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6841 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6901 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6961 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7021 cagggttatt gtctcatgtc cgcgcgtt
//
