LOCUS       sequence-169612-        6503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Venus tagged Cdc-42 sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6503)
  AUTHORS   O'Neill PR, Kalyanaraman V, Gautam N
  TITLE     Subcellular optogenetic activation of Cdc42 controls local and 
            distal signaling to drive immune cell migration.
  JOURNAL   Mol Biol Cell. 2016 May 1;27(9):1442-50. doi: 
            10.1091/mbc.E15-12-0832. Epub 2016 Mar 3.
  PUBMED    26941336
REFERENCE   2  (bases 1 to 6503)
  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: 
            "10.1091/mbc.E15-12-0832"; journalName: "Mol Biol Cell"; date: 
            "2016-05-1- 1"; volume: "27"; issue: "9"; pages: "1442-50"
COMMENT     Sequence Label: Venus wGBD
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(292..312)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        297..626
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      477..612
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     539..558
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             693..1487
                     /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(747..766)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1357..1376
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1661..1782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1698..1717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1752..1771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1831..1847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1831..1847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1844..1866)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1855..1871
                     /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(1879..1885,1886..1903,1904..1909))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1924..1945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2062..2079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2233..2821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2313..2332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2992..3783,3784..3852))
                     /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     3615..3634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3853..3957)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4032..4051)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4223..4602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4603..4806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4757..4777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4851..4870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4851..4869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(4905..4907,4908..4910,4911..5621)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4950..4971)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5211..5230)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5558..5579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6082..6099)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6088..6312
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6358..6503,1..283)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6445..6464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
       61 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      121 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
      181 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
      241 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
      301 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      361 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      421 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      481 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      541 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
      601 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
      661 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
      721 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
      781 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
      841 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
      901 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
      961 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     1021 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     1081 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     1141 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     1201 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     1261 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     1321 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     1381 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     1441 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     1501 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     1561 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     1621 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     1681 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1741 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     1801 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     1861 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1921 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1981 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2041 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2101 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2161 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2221 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2281 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2341 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2401 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2461 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2521 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2581 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2641 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2701 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2761 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2821 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2881 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2941 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3001 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3061 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3121 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3181 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3241 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3301 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3361 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3421 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3481 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3541 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     3601 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     3661 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     3721 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     3781 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     3841 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     3901 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     3961 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     4021 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     4081 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     4141 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     4201 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     4261 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     4321 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     4381 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4441 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     4501 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     4561 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     4621 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     4681 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     4741 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     4801 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     4861 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttatggtg agcaagggcg
     4921 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4981 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5041 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     5101 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     5161 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     5221 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     5281 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     5341 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     5401 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     5461 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     5521 ccaaactgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     5581 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggtacccct agcccagctg
     5641 ataagaaacg ctcagggaag aagaagatca gcaaagctga tattggtgca cccagtggat
     5701 tcaagcatgt cagccacgtg gggtgggacc cccagaatgg atttgacgtg aacaacctcg
     5761 acccagatct gcggagtctg ttctccaggg caggaatcag cgaggcccag ctcaccgacg
     5821 ccgagacctc taaacttatc tacgacttca ttgaggacca gggtgggctg gaggctgtgc
     5881 ggcaggagat gaggcgccag gagttctaga gaattccgat ttccaaagcg ggaaaggaaa
     5941 tcaaggagta cgtggaggcc caagcaggaa acgatccttt tctcaaaggc atccctgagg
     6001 acaagaatcc cttcaaggag aaaggtggct gtctgataag ctgactcgag tctagagggc
     6061 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6121 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6181 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6241 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     6301 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     6361 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6421 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6481 tcgccggctt tccccgtcaa gct
//
