LOCUS       sequence-138865-        6768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of photoactivatable Rac1.
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-mCerulean-PA-Rac1
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(158..745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(237..256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(758..1549,1550..1618))
                     /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     1381..1400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     1861..2688
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             1861..1985
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(2023..2628)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        2756..3059
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3061..3263
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3214..3234
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3466..3488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3550..3569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     3611..3630
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3611..3629
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3630..3654
                     /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        3670..3779
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             3800..3817
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(3824..3826,3827..4537)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3866..3887)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4127..4146)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4474..4495
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4556..4984
                     /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             5591..5623
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             5630..5653
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(5743..5762)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5808..5863
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5862..5881)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      5951..5998
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     6011..6716
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           6099..6471
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(6128..6716)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(6725..6758)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
       61 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgccccctga
      181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      421 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      481 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      541 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
      601 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
      661 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
      721 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgtta ccaatgctta atcagtgagg
      781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
      841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
      901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
      961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     1021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     1081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     1141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     1201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     1261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     1321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     1381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     1441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     1501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     1561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     1621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg tccgcgcgtt
     1681 tcctgcatct tttaatcaaa tcccaagatg tgtataaacg cgccggtatg tacaggaaga
     1741 ggtttatact aaactgttac attgcaaacg tggtttcgtg tgccaagtgt gaaaaccgat
     1801 gtttaatcaa ggctctgacg catttctaca accacgactc caagtgtgtg ggtgaagtca
     1861 tgcatctttt aatcaaatcc caagatgtgt ataaaccacc aaactgccaa aaaatgaaaa
     1921 ctgtcgacaa gctctgtccg tttgctggca actgcaaggg tctcaatcct atttgtaatt
     1981 attgaataat aaaacaatta taaatgtcaa atttgttttt tattaacgat acaaaccaaa
     2041 cgcaacaaga acatttgtag tattatctat aattgaaaac gcgtagttat aatcgctgag
     2101 gtaatattta aaatcatttt caaatgattc acagttaatt tgcgacaata taattttatt
     2161 ttcacataaa ctagacgcct tgtcgtcttc ttcttcgtat tccttctctt tttcattttt
     2221 ctcttcataa aaattaacat agttattatc gtatccatat atgtatctat cgtatagagt
     2281 aaattttttg ttgtcataaa tatatatgtc ttttttaatg gggtgtatag taccgctgcg
     2341 catagttttt ctgtaattta caacagtgct attttctggt agttcttcgg agtgtgttgc
     2401 tttaattatt aaatttatat aatcaatgaa tttgggatcg tcggttttgt acaatatgtt
     2461 gccggcatag tacgcagctt cttctagttc aattacacca ttttttagca gcaccggatt
     2521 aacataactt tccaaaatgt tgtacgaacc gttaaacaaa aacagttcac ctcccttttc
     2581 tatactattg tctgcgagca gttgtttgtt gttaaaaata acagccattg taatgagacg
     2641 cacaaactaa tatcacaaac tggaaatgtc tatcaatata tagttgctga tggccggcct
     2701 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2761 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2821 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2881 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc
     2941 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     3001 ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     3061 tgatgcggtt ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc
     3121 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     3181 ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt
     3241 gggaggtcta tataagcaga cgtcgtttag tgaaccgtca gatcactaga tgctttattg
     3301 cggtagttta tcacagttaa attgctaacg ccagtctcga acttaacgtg cagaagttgg
     3361 tcgtgaggca ctgggcaggt aagtatcggg ccctttgtgc ggggggagcg gctcggggct
     3421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     3481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     3541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
     3601 ggaccgaaat taatacgact cactataggg gaattgtgag cggataacaa ttccccggag
     3661 ttaatccggg acctttaatt caacccaaca caatatatta tagttaaata agaattatta
     3721 tcaaatcatt tgtatattaa ttaaaatact atactgtaaa ttacatttta tttacaatca
     3781 aaggagatat accatggcac accatcacca ccatcacggt tctgtgagca agggcgagga
     3841 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3901 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3961 catcagcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg
     4021 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4081 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4141 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4201 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat
     4261 cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     4321 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4381 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     4441 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4501 cgccgggatc actctcggca tggacgagct gtacaagggt tctggaagtg gatccttggc
     4561 tactacactt gaacgtattg agaagaactt tgtcattact gacccaagat tgccagataa
     4621 tcccattata ttcgcgtccg atagtttctt gcagttgaca gaatatagcc gtgaagaaat
     4681 tttgggaaga aactgcaggt ttctacaagg tcctgaaact gatcgcgcga cagtgagaaa
     4741 aattagagat gccatagata accaaacaga ggtcactgtt cagctgatta attatacaaa
     4801 gagtggtaaa aagttctgga acctctttca cttgcagcct atgcgagatc agaagggaga
     4861 tgtccagtac tttattgggg ttcagttgga tggaactgag catgtccgag atgctgccga
     4921 gagagaggga gtcatgctga ttaagaaaac tgcagaaaat attgatgagg cggcaaaaga
     4981 acttatcaag tgtgtggtgg tgggagacgg agctgtaggt aaaacttgcc tactgatcag
     5041 ttacacaacc aatgcatttc ctggagaata tatccctact gtctttgaca attattctgc
     5101 caatgttatg gtagatggaa aaccggtgaa tctgggctta tgggatacag ctggactaga
     5161 agattatgac agattacgcc ccctatccta tccgcaaaca gatgtgttct taatttgctt
     5221 ttcccttgtg agtcctgcat cattccacca cgtccgtgca aagtggtatc ctgaggtgcg
     5281 gcaccactgt cccaacactc ccatcatcct agtgggaact aaacttgatc ttagggatga
     5341 taaagacacg atcgagaaac tgaaggagaa gaagctgact cccatcacct atccgcaggg
     5401 tctagccatg gctaaggaga ttggtgctgt aaaatacctg gagtgctcgg cgctcacaca
     5461 gcgaggcctc aagacagtgt ttgacgaagc gatccgagca gtcctctgcc cgcctcccgt
     5521 gaagaagagg aagagaaaat gcctgctgtt gtaaaagctt gcggccgcac agctgtatac
     5581 acgtgcaagc cagccagaac tcgccccgga agaccccgag gatctcgagc accaccatca
     5641 ccatcaccat cactaagtga ttaacctcag gtgcaggctg cctatcagaa ggtggtggct
     5701 ggtgtggcca atgccctggc tcacaaatac cactgagatc gatctttttc cctctgccaa
     5761 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     5821 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     5881 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     5941 ccatatgtaa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     6001 gaaagcatgc ggaggaaatt ctccttgaag tttccctggt gttcaaagta aaggagtttg
     6061 caccagacgc acctctgttc actggtccgg cgtattaaaa cacgatacat tgttattagt
     6121 acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt tgtacgtatt
     6181 ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa atcaaatgat
     6241 tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg taaaataggt
     6301 ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta tctaatggat
     6361 tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct ttgtcgatat
     6421 tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg
     6481 tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg ttaaatagag
     6541 cttggacata tttaacatcg ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc
     6601 aaccctcgtc gttagaagtt gcttccgaag acgattttgc catagccaca cgacgcctat
     6661 taattgtgtc ggctaacacg tccgcgatca aatttgtagt tgagcttttt ggaattgcga
     6721 tcgcataact tcgtatagca tacattatac gaagttataa gctcggaa
//
