LOCUS       sequence-204194-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active Cdc42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  AUTHORS   Petrie RJ, Gavara N, Chadwick RS, Yamada KM
  TITLE     Nonpolarized signaling reveals two distinct modes of 3D cell 
            migration.
  JOURNAL   J Cell Biol. 2012 Apr 30;197(3):439-55. doi: 10.1083/jcb.201201124.
  PUBMED    22547408
REFERENCE   2  (bases 1 to 5283)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2012-04-30- 30"; volume: "197"; 
            issue: "3"; pages: "439-55"
COMMENT     Sequence Label: pECFP-Cdc42Q61L
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        790..1093
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1094..1297
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1248..1268
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1336..1345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1342..1344,1345..1347,1348..2058)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1387..1408)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1648..1667)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1995..2016
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(2095..2274,2275..2277,2278..2670)
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase Cdc42"
                     /label=Cdc42 (Q61L)
                     /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVM
                     IGGEPYTLGLFDTAGLEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH
                     CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK
                     GLKNVFDEAILAALEPPEPKKSRRCVLL"
     polyA_signal    2800..2921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2837..2856)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2891..2910
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2928..3383)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3065..3086)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3277..3296
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3410..3514
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3516..3873
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3516..3707
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3539..3559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3724..3859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3908..4702
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3908..4702
                     /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(3962..3981)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4572..4591
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4890..4909)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4934..4981
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttaggcca ccacttcaag aactctgtat caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      781 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      841 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      901 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      961 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1021 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     1081 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     1141 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1201 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1261 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
     1321 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1381 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1441 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1501 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
     1561 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1621 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1681 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1741 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
     1801 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1861 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1921 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1981 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2041 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttccatgcag
     2101 acaattaagt gtgttgttgt gggcgatggt gccgttggta aaacatgtct cctgatatcc
     2161 tacacaacaa acaaatttcc atctgagtat gtaccgactg tttttgacaa ctatgcagtc
     2221 acggttatga ttggtggaga gccatatacc cttggacttt ttgatactgc agggctagag
     2281 gattatgaca gattacgacc gctgagttat ccacaaacag atgtatttct agtctgtttt
     2341 tcagtggtct ctccatcctc atttgaaaat gtgaaggaaa agtgggtgcc tgagataact
     2401 caccactgtc caaagactcc tttcttgctt gttgggaccc aaattgatct ccgagatgac
     2461 ccctctacta tcgagaaact tgccaagaac aaacagaagc ctatcactcc agagactgct
     2521 gaaaagctgg cccgtgacct gaaggcggtc aagtatgtgg agtgttctgc actcacacag
     2581 aaaggcctaa agaatgtatt tgacgaagca atattggctg ccctggagcc tccagaaccg
     2641 aagaagagcc gcaggtgtgt gctgctatga ggatccaccg gatctagata actgatcata
     2701 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2761 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2821 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2881 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2941 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3001 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3061 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3121 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3181 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3241 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3301 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3361 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3421 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3481 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3541 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3601 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3661 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3721 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3781 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3841 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3901 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3961 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4021 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4081 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4141 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4201 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4261 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4321 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4381 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4441 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4501 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4561 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4621 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4681 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4861 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4921 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4981 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5041 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5101 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5161 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5221 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5281 gac
//
