LOCUS       sequence-138512-        6768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of photoactivatable Rac1 (dominant-negative mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6768)
  AUTHORS   Wu YI, Frey D, Lungu OI, Jaehrig A, Schlichting I, Kuhlman B, Hahn 
            KM
  TITLE     A genetically encoded photoactivatable Rac controls the motility of 
            living cells.
  JOURNAL   Nature. 2009 Sep 3. 461(7260):104-8.
  PUBMED    19693014
REFERENCE   2  (bases 1 to 6768)
  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: "Nature. 
            2009 Sep 3. 461(7260):104-8."
COMMENT     Sequence Label: pTriEx-mVenus-PA-Rac1-T17N
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1..706
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             1..589
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     3'UTR           246..618
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     terminator      complement(719..766)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     836..855
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    854..909
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     955..974
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1064..1087)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(1094..1126)
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             complement(1733..2161)
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             complement(join(2180..2890,2891..2893))
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2222..2243)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2571..2590
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2830..2851
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2900..2917)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2938..3047)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     protein_bind    3063..3087
                     /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)."
     primer_bind     complement(3087..3106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3088..3106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3148..3167)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3229..3251)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3454..3656)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3483..3503)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3658..3961
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_recomb     4029..4856
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4089..4694
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     CDS             complement(4732..4856)
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             join(5099..5167,5168..5959)
                     /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     complement(5317..5336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5972..6559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6461..6480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6727..6760
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aattccaaaa agctcaacta caaatttgat cgcggacgtg ttagccgaca caattaatag
       61 gcgtcgtgtg gctatggcaa aatcgtcttc ggaagcaact tctaacgacg agggttggga
      121 cgacgacgat aatcggccta ataaagctaa cacgcccgat gttaaatatg tccaagctct
      181 atttaacgtg tttacgtcga gtcaattgta cactaacgac agtgatgaaa gaaatacaaa
      241 agcgcataat attttgaacg acgtcgaacc tttattacaa aacaaaacac aaacgaatat
      301 cgacaaagct agattgctgc tacaagattt ggcaagtttt gtggcgttga gcgaaaatcc
      361 attagatagt ccagccatcg gttcggaaaa acaacccttg tttgaaacta atcgaaacct
      421 attttacaaa tctattgagg atttaatatt taaattcaga tataaagacg ctgaaaatca
      481 tttgattttc gctctaacat accaccctaa agattataaa tttaatgaat tattaaaata
      541 cgtacaacaa ttgtctgtaa atcaacaacg cacagaatct agcgcttaat aaatgtacta
      601 ataacaatgt atcgtgtttt aatacgccgg accagtgaac agaggtgcgt ctggtgcaaa
      661 ctcctttact ttgaacacca gggaaacttc aaggagaatt tcctccgcat gctttcagca
      721 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttac atatgggcat
      781 atgttgccaa actctaaacc aaatactcat tctgatgttt taaatgattt gccctcccat
      841 atgtccttcc gagtgagaga cacaaaaaat tccaacacac tattgcaatg aaaataaatt
      901 tcctttatta gccagaagtc agatgctcaa ggggcttcat gatgtcccca taatttttgg
      961 cagagggaaa aagatcgatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc
     1021 accaccttct gataggcagc ctgcacctga ggttaatcac ttagtgatgg tgatggtgat
     1081 ggtggtgctc gagatcctcg gggtcttccg gggcgagttc tggctggctt gcacgtgtat
     1141 acagctgtgc ggccgcaagc ttttacaaca gcaggcattt tctcttcctc ttcttcacgg
     1201 gaggcgggca gaggactgct cggatcgctt cgtcaaacac tgtcttgagg cctcgctgtg
     1261 tgagcgccga gcactccagg tattttacag caccaatctc cttagccatt gctagaccct
     1321 gcggataggt gatgggagtc agcttcttct ccttcagttt ctcgatcgtg tctttatcat
     1381 ccctaagatc aagtttagtt cccactagga tgatgggagt gttgggacag tggtgccgca
     1441 cctcaggata ccactttgca cggacatttt caaatgatgc aggactcaca agggaaaagc
     1501 aaattaagaa cacatctgtt tgcggatagg atagggggcg taatctgtca taatcttctt
     1561 gtccagctgt atcccataag cccagattca ccggttttcc atctaccata acattggcag
     1621 aataattgtc aaagacagta gggatatatt ctccaggaaa tgcattggtt gtgtaactga
     1681 tcagtaggca atttttacct acagctccgt ctcccaccac cacacacttg ataagttctt
     1741 ttgccgcctc atcaatattt tctgcagttt tcttaatcag catgactccc tctctctcgg
     1801 cagcatctcg gacatgctca gttccatcca actgaacccc aataaagtac tggacatctc
     1861 ccttctgatc tcgcataggc tgcaagtgaa agaggttcca gaacttttta ccactctttg
     1921 tataattaat cagctgaaca gtgacctctg tttggttatc tatggcatct ctaatttttc
     1981 tcactgtcgc gcgatcagtt tcaggacctt gtagaaacct gcagtttctt cccaaaattt
     2041 cttcacggct atattctgtc aactgcaaga aactatcgga cgcgaatata atgggattat
     2101 ctggcaatct tgggtcagta atgacaaagt tcttctcaat acgttcaagt gtagtagcca
     2161 aggatccact tccagaaccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     2221 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagtttgg
     2281 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     2341 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
     2401 agtttgcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     2461 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     2521 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     2581 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     2641 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc aggccgtagc
     2701 cgagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg ctgatgaact
     2761 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     2821 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     2881 cgcccttgct cacagaaccg tgatggtggt gatggtgtgc catggtatat ctcctttgat
     2941 tgtaaataaa atgtaattta cagtatagta ttttaattaa tatacaaatg atttgataat
     3001 aattcttatt taactataat atattgtgtt gggttgaatt aaaggtcccg gattaactcc
     3061 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cggtccgatc
     3121 caattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca ggagctgtag
     3181 gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg tcacacgcca
     3241 gaagccgaac cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc gcggacagcc
     3301 ccgagccgct ccccccgcac aaagggcccg atacttacct gcccagtgcc tcacgaccaa
     3361 cttctgcacg ttaagttcga gactggcgtt agcaatttaa ctgtgataaa ctaccgcaat
     3421 aaagcatcta gtgatctgac ggttcactaa acgacgtctg cttatataga cctcccaccg
     3481 tacacgccta ccgcccattt gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc
     3541 ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa
     3601 tccccgtgag tcaaaccgct atccacgccc attggtgtac tgccaaaacc gcatcagcat
     3661 ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat
     3721 gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag ggggcggact
     3781 tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca
     3841 ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt
     3901 caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac
     3961 gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact attaataggc
     4021 cggccatcag caactatata ttgatagaca tttccagttt gtgatattag tttgtgcgtc
     4081 tcattacaat ggctgttatt tttaacaaca aacaactgct cgcagacaat agtatagaaa
     4141 agggaggtga actgtttttg tttaacggtt cgtacaacat tttggaaagt tatgttaatc
     4201 cggtgctgct aaaaaatggt gtaattgaac tagaagaagc tgcgtactat gccggcaaca
     4261 tattgtacaa aaccgacgat cccaaattca ttgattatat aaatttaata attaaagcaa
     4321 cacactccga agaactacca gaaaatagca ctgttgtaaa ttacagaaaa actatgcgca
     4381 gcggtactat acaccccatt aaaaaagaca tatatattta tgacaacaaa aaatttactc
     4441 tatacgatag atacatatat ggatacgata ataactatgt taatttttat gaagagaaaa
     4501 atgaaaaaga gaaggaatac gaagaagaag acgacaaggc gtctagttta tgtgaaaata
     4561 aaattatatt gtcgcaaatt aactgtgaat catttgaaaa tgattttaaa tattacctca
     4621 gcgattataa ctacgcgttt tcaattatag ataatactac aaatgttctt gttgcgtttg
     4681 gtttgtatcg ttaataaaaa acaaatttga catttataat tgttttatta ttcaataatt
     4741 acaaatagga ttgagaccct tgcagttgcc agcaaacgga cagagcttgt cgacagtttt
     4801 cattttttgg cagtttggtg gtttatacac atcttgggat ttgattaaaa gatgcatgac
     4861 ttcacccaca cacttggagt cgtggttgta gaaatgcgtc agagccttga ttaaacatcg
     4921 gttttcacac ttggcacacg aaaccacgtt tgcaatgtaa cagtttagta taaacctctt
     4981 cctgtacata ccggcgcgtt tatacacatc ttgggatttg attaaaagat gcaggaaacg
     5041 cgcggacatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5101 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5161 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5221 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5281 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5341 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5401 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5461 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5521 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5581 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5641 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5701 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5761 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5821 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5881 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5941 actgattaag cattggtaac aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6541 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6601 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     6661 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgttcc
     6721 gagcttataa cttcgtataa tgtatgctat acgaagttat gcgatcgc
//
