LOCUS       Exported                7321 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Also known as optoGEF-RhoA, encodes for a fusion between 
            CRY2(1-498)-mCherry and the DHPH domain of ARHGEF11.
ACCESSION   .
VERSION     .
KEYWORDS    ARHGEF11(DHPH)-CRY2-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7321)
  AUTHORS   Valon L, Marin-Llaurado A, Wyatt T, Charras G, Trepat X
  TITLE     Optogenetic control of cellular forces and mechanotransduction.
  JOURNAL   Nat Commun. 2017 Feb 10;8:14396. doi: 10.1038/ncomms14396.
  PUBMED    28186127
REFERENCE   2  (bases 1 to 7321)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7321
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1751..3244
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      3263..3272
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3269..3979
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3414..3432)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3682..3701
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4102..4223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4193..4212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4230..4685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4367..4388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4579..4598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4712..4816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4818..5175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4841..4861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5026..5161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5088..5107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5210..6004
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5264..5283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5874..5893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6192..6211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6236..6283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6612..7200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7101..7120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagca tgctgaccca
      601 gagagaaatt gataggcagg aagtgattaa cgagctgttt gtgaccgagg caagccatct
      661 gagaaccctg agagtgctgg acctgatctt ctaccagagg atgaagaaag agaacctgat
      721 gccccgggag gaactggcta gactgtttcc aaacctgccc gagctgatcg aaattcacaa
      781 ttcctggtgc gaagcaatga agaaactgcg cgaggaagga cctatcatta aggagatttc
      841 tgacctgatg ctggctaggt tcgatggccc agcacgcgag gaactgcagc aggtggccgc
      901 tcagttttgc tcataccaga gcatcgccct ggaactgatt aagactaaac agagaaagga
      961 gtctaggttc cagctgttta tgcaggaggc tgaaagtcac ccacagtgtc ggagactgca
     1021 gctgcgcgac ctgatcatta gcgaaatgca gcgactgaca aagtatcccc tgctgctgga
     1081 aagcatcatc aagcacactg agggcgggac ctccgagcat gaaaagctgt gcagggcccg
     1141 cgatcagtgt cgggagatcc tgaagtacgt gaacgaggct gtcaaacaga ccgaaaatcg
     1201 acatcggctg gagggctatc agaaaagact ggacgctaca gcactggaaa gggccagtaa
     1261 ccccctggca gccgagttca agtcactgga tctgaccaca cggaaaatga tccacgaggg
     1321 gcctctgacc tggagaatta gtaaggacaa aacactggat ctgcatgtgc tgctgctgga
     1381 agacctgctg gtcctgctgc agaagcagga tgagaaactg ctgctgaagt gccacagcaa
     1441 aactgcagtg ggcagctccg actccaagca gaccttttct cctgtcctga aactgaatgc
     1501 cgtgctgatc cgatcagtcg caacagataa gcgggccttc tttatcattt gtactagcaa
     1561 actggggccc cctcagatct acgaactggt ggccctgaca tctagtgata agaacacttg
     1621 gatggagctg ctggaggaag ccgtgcgcaa tgctacccga catccaggag agttctccgc
     1681 aggggggtca gcaggggggt ccgcaggggg gtccgcaggg gggtcagcag gaggggctct
     1741 cgaggccacc atgaagatgg acaaaaagac tatagtttgg tttagaagag acctaaggat
     1801 tgaggataat cctgcattag cagcagctgc tcacgaagga tctgtttttc ctgtcttcat
     1861 ttggtgtcct gaagaagaag gacagtttta tcctggaaga gcttcaagat ggtggatgaa
     1921 acaatcactt gctcacttat ctcaatcctt gaaggctctt ggatctgacc tcactttaat
     1981 caaaacccac aacacgattt cagcgatctt ggattgtatc cgcgttaccg gtgctacaaa
     2041 agtcgtcttt aaccacctct atgatcctgt ttcgttagtt cgggaccata ccgtaaagga
     2101 gaagctggtg gaacgtggga tctctgtgca aagctacaat ggagatctat tgtatgaacc
     2161 gtgggagata tactgcgaaa agggcaaacc ttttacgagt ttcaattctt actggaagaa
     2221 atgcttagat atgtcgattg aatccgttat gcttcctcct ccttggcggt tgatgccaat
     2281 aactgcagcg gctgaagcga tttgggcgtg ttcgattgaa gaactagggc tggagaatga
     2341 ggccgagaaa ccgagcaatg cgttgttaac tagagcttgg tctccaggat ggagcaatgc
     2401 tgataagtta ctaaatgagt tcatcgagaa gcagttgata gattatgcaa agaacagcaa
     2461 gaaagttgtt gggaattcta cttcactact ttctccgtat ctccatttcg gggaaataag
     2521 cgtcagacac gttttccagt gtgcccggat gaaacaaatt atatgggcaa gagataagaa
     2581 cagtgaagga gaagaaagtg cagatctttt tcttagggga atcggtttaa gagagtattc
     2641 tcggtatata tgtttcaact tcccgtttac tcacgagcaa tcgttgttga gtcatcttcg
     2701 gtttttccct tgggatgctg atgttgataa gttcaaggcc tggagacaag gcaggaccgg
     2761 ttatccgttg gtggatgccg gaatgagaga gctttgggct accggatgga tgcataacag
     2821 aataagagtg attgtttcaa gctttgctgt gaagtttctt ctccttccat ggaaatgggg
     2881 aatgaagtat ttctgggata cacttttgga tgctgatttg gaatgtgaca tccttggctg
     2941 gcagtatatc tctgggagta tccccgatgg ccacgagctt gatcgcttgg acaatcccgc
     3001 gttacaaggc gccaaatatg acccagaagg tgagtacata aggcaatggc ttcccgagct
     3061 tgcgagattg ccaactgaat ggatccatca tccatgggac gctcctttaa ccgtactcaa
     3121 agcttctggt gtggaactcg gaacaaacta tgcgaaaccc attgtagaca tcgacacagc
     3181 tcgtgagcta ctagctaaag ctatttcaag aacccgtgaa gcacagatca tgatcggagc
     3241 agcagcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
     3301 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     3361 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3421 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     3481 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     3541 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3601 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     3661 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3721 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     3781 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     3841 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     3901 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     3961 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4201 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     4261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4681 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4741 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4801 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4921 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5041 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5101 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5161 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     5221 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     5281 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5341 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5401 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5461 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5521 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5581 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5641 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5701 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5761 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5821 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5881 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5941 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6001 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6061 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6121 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6181 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     6241 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6301 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6361 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6421 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6481 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6541 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6601 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6661 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6721 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6781 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6841 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6901 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6961 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7021 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7081 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7141 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7201 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7261 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     7321 t
//
