LOCUS       sequence-55893-e        7425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses caged Cdc42 GEF intersectin: Intersectin Dbl homology (DH)
            domain fused to Dronpa145N at C terminus and at N terminus, for 
            optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7425)
  AUTHORS   Zhou XX, Chung HK, Lam AJ, Lin MZ
  TITLE     Optical control of protein activity by fluorescent protein domains.
  JOURNAL   Science. 2012 Nov 9;338(6108):810-4. doi: 10.1126/science.1226854.
  PUBMED    23139335
REFERENCE   2  (bases 1 to 7425)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2012-11-9- 9"; volume: "338"; issue: 
            "6108"; pages: "810-4"
COMMENT     Sequence Label: pcDNA3-N-I-N-CAAX
FEATURES             Location/Qualifiers
     source          1..7425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2977..2995)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2978..2995)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3015..3032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3021..3245
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3291..3719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3378..3397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3588..3609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3728..3748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3733..4063
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3914..4049
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3976..3995
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4130..4924
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4184..4203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4794..4813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5098..5219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5135..5154)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5189..5208
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5268..5284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5268..5284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5281..5303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5292..5308
                     /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        complement(join(5316..5322,5323..5340,5341..5346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5499..5516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6429..7220,7221..7289))
                     /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     7052..7071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgagc
      901 gtgatcaagc ccgacatgaa aatcaagctg aggatggagg gtgccgtgaa cggtcacccg
      961 tttgcgatcg agggcgtggg actgggcaag ccgttcgagg gaaagcaaag catggacctc
     1021 aaggtgaagg agggcggacc gctgcccttc gcctacgaca ttctgaccac tgtcttctgc
     1081 tacggtaatc gcgtcttcgc aaagtatccc gaaaacatcg tggattactt taagcagagc
     1141 ttcccggagg gttactcctg ggagcgatcc atgaactacg aggacggagg gatctgcaac
     1201 gcgacgaacg atataacctt ggacggcgac tgctacattt acgaaattcg gttcgacggc
     1261 gtgaatttcc cagcgaatgg ccctgtaatg caaaaacgca cggtaaagtg ggagcccagc
     1321 accgagaacc tgtacgtccg cgacggagtc ctgaagggag acgtgaacat ggcgctttca
     1381 ttggaaggtg gcgggcacta cagatgcgat ttcaagacga cctataaggc gaaaaaggtg
     1441 gtccaactgc cggactacca tttcgtcgac caccacatcg agatcaagag ccacgacaaa
     1501 gactatagca acgtgaatct ccacgagcat gccgaggccc acagcgagct gcccaggcag
     1561 gccaagggcc ttaagggatc catgactgaa aggaagaggc agggctacat ccacgagctg
     1621 attgtcacgg aagagaatta cgtgaacgac ctgcagctgg tgaccgagat ctttcagaag
     1681 ccgcttatgg agagtgaact tttgacggaa aaagaggtcg ccatgatctt tgtgaactgg
     1741 aaggagctta taatgtgcaa catcaagctg ctgaaggccc ttagggtgcg gaagaagatg
     1801 tctggtgaga aaatgccggt caaaatgatc ggcgacatcc tgtctgccca gctcccccac
     1861 atgcagccct acatcaggtt ctgcagcagg caactcaacg gtgcggccct gatccagcag
     1921 aagaccgacg aggcacccga cttcaaagag ttcgtgaaac ggctggagat ggaccccagg
     1981 tgcaagggaa tgccgctctc atccttcatc ctcaagccga tgcagagggt tacccgatac
     2041 cccctgatta tcaagaatat cctggagaac acccccgaga accaccccga ccacagccac
     2101 ctgaagcacg cgctggaaaa ggcggaggag ttgtgcagcc aggttaacga gggcgtgagg
     2161 gagaaggaag gcagtagcgg aagcagtggt tcttcaggat catccgaatt cggcatgagc
     2221 gtgatcaagc ccgacatgaa aatcaagctg aggatggagg gtgccgtgaa cggtcacccg
     2281 tttgcgatcg agggcgtggg actgggcaag ccgttcgagg gaaagcaaag catggacctc
     2341 aaggtgaagg agggcggacc gctgcccttc gcctacgaca ttctgaccac tgtcttctgc
     2401 tacggtaatc gcgtcttcgc aaagtatccc gaaaacatcg tggattactt taagcagagc
     2461 ttcccggagg gttactcctg ggagcgatcc atgaactacg aggacggagg gatctgcaac
     2521 gcgacgaacg atataacctt ggacggcgac tgctacattt acgaaattcg gttcgacggc
     2581 gtgaatttcc cagcgaatgg ccctgtaatg caaaaacgca cggtaaagtg ggagcccagc
     2641 accgagaacc tgtacgtccg cgacggagtc ctgaagggag acgtgaacat ggcgctttca
     2701 ttggaaggtg gcgggcacta cagatgcgat ttcaagacga cctataaggc gaaaaaggtg
     2761 gtccaactgc cggactacca tttcgtcgac caccacatcg agatcaagag ccacgacaaa
     2821 gactatagca acgtgaatct ccacgagcat gccgaggccc acagcgagct gcccaggcag
     2881 gccaagaaga tgagcaagga cggcaagaag aaaaagaaga agtccaagac taagtgcgtg
     2941 atcatgtaac tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3001 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3061 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3121 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3181 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3241 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3421 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     3481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3661 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3721 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     3781 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3841 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3901 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3961 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4021 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4081 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4141 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4201 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4261 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4321 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4381 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4441 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4501 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4561 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4621 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4681 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4741 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4801 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4861 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4921 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4981 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5041 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5101 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5161 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5221 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5281 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5341 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5401 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
