LOCUS       sequence-240702-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses caged Cdc42 GEF intersectin: Intersectin Dbl homology (DH)
            domain fused to wild-type Dronpa145K at C terminus and Dronpa145N at
            N terminus, for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  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 7424)
  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-K-I-N-CAAX
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        155..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        535..738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     689..709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     783..802
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        783..801
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2897..2915)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2898..2915)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2935..2952)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2941..3165
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3211..3639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3508..3529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3648..3668)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3653..3982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3833..3968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3895..3914
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4049..4843
                     /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(4103..4122)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4713..4732
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5017..5138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5054..5073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5108..5127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5187..5203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5187..5203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5200..5222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5211..5227
                     /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(5235..5241,5242..5259,5260..5265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5280..5301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5418..5435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5589..6177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5669..5688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6348..7139,7140..7208))
                     /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     6971..6990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7209..7313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7388..7407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
       61 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      121 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      721 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      781 attaatacga ctcactatag ggagacccaa gcttatgagc gtgatcaagc ccgacatgaa
      841 aatcaagctg aggatggagg gtgccgtgaa cggtcacccg tttgcgatcg agggcgtggg
      901 actgggcaag ccgttcgagg gaaagcaaag catggacctc aaggtgaagg agggcggacc
      961 gctgcccttc gcctacgaca ttctgaccac tgtcttctgc tacggtaatc gcgtcttcgc
     1021 aaagtatccc gaaaacatcg tggattactt taagcagagc ttcccggagg gttactcctg
     1081 ggagcgatcc atgaactacg aggacggagg gatctgcaac gcgacgaacg atataacctt
     1141 ggacggcgac tgctacattt acgaaattcg gttcgacggc gtgaatttcc cagcgaatgg
     1201 ccctgtaatg caaaaacgca cggtaaagtg ggagcccagc accgagaagc tgtacgtccg
     1261 cgacggagtc ctgaagggag acgtgaacat ggcgctttca ttggaaggtg gcgggcacta
     1321 cagatgcgat ttcaagacga cctataaggc gaaaaaggtg gtccaactgc cggactacca
     1381 tttcgtcgac caccacatcg agatcaagag ccacgacaaa gactatagca acgtgaatct
     1441 ccacgagcat gccgaggccc acagcgagct gcccaggcag gccaagggcc ttaagggatc
     1501 catgactgaa aggaagaggc agggctacat ccacgagctg attgtcacgg aagagaatta
     1561 cgtgaacgac ctgcagctgg tgaccgagat ctttcagaag ccgcttatgg agagtgaact
     1621 tttgacggaa aaagaggtcg ccatgatctt tgtgaactgg aaggagctta taatgtgcaa
     1681 catcaagctg ctgaaggccc ttagggtgcg gaagaagatg tctggtgaga aaatgccggt
     1741 caaaatgatc ggcgacatcc tgtctgccca gctcccccac atgcagccct acatcaggtt
     1801 ctgcagcagg caactcaacg gtgcggccct gatccagcag aagaccgacg aggcacccga
     1861 cttcaaagag ttcgtgaaac ggctggagat ggaccccagg tgcaagggaa tgccgctctc
     1921 atccttcatc ctcaagccga tgcagagggt tacccgatac cccctgatta tcaagaatat
     1981 cctggagaac acccccgaga accaccccga ccacagccac ctgaagcacg cgctggaaaa
     2041 ggcggaggag ttgtgcagcc aggttaacga gggcgtgagg gagaaggaag gcagtagcgg
     2101 aagcagtggt tcttcaggat catccgaatt cggcatgagc gtgatcaagc ccgacatgaa
     2161 aatcaagctg aggatggagg gtgccgtgaa cggtcacccg tttgcgatcg agggcgtggg
     2221 actgggcaag ccgttcgagg gaaagcaaag catggacctc aaggtgaagg agggcggacc
     2281 gctgcccttc gcctacgaca ttctgaccac tgtcttctgc tacggtaatc gcgtcttcgc
     2341 aaagtatccc gaaaacatcg tggattactt taagcagagc ttcccggagg gttactcctg
     2401 ggagcgatcc atgaactacg aggacggagg gatctgcaac gcgacgaacg atataacctt
     2461 ggacggcgac tgctacattt acgaaattcg gttcgacggc gtgaatttcc cagcgaatgg
     2521 ccctgtaatg caaaaacgca cggtaaagtg ggagcccagc accgagaacc tgtacgtccg
     2581 cgacggagtc ctgaagggag acgtgaacat ggcgctttca ttggaaggtg gcgggcacta
     2641 cagatgcgat ttcaagacga cctataaggc gaaaaaggtg gtccaactgc cggactacca
     2701 tttcgtcgac caccacatcg agatcaagag ccacgacaaa gactatagca acgtgaatct
     2761 ccacgagcat gccgaggccc acagcgagct gcccaggcag gccaagaaga tgagcaagga
     2821 cggcaagaag aaaaagaaga agtccaagac taagtgcgtg atcatgtaac tcgagcatgc
     2881 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3121 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     3181 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3661 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3721 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3841 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3901 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3961 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4021 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4081 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4141 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4201 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4261 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4321 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4381 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4441 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4501 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4561 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4621 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4681 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4741 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4801 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4861 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4921 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4981 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     5041 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5101 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     5161 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5221 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5281 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5341 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5401 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5461 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5521 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5761 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6001 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6241 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6301 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6361 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6421 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6481 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6541 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6601 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6661 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6721 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6781 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6841 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6901 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6961 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7021 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7081 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7141 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7201 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7261 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7321 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     7381 ctctcagtac aatctgctct gatgccgcat agttaagcca gtat
//
