LOCUS       sequence-204197-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control for FRET biosensor analysis - cannot bind downstream 
            effectors.
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-Cdc42-Y40C
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     511..530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        785..1088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1089..1292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1243..1263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1331..1340
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1337..1339,1340..1342,1343..2053)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1382..1403)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1643..1662)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1990..2011
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2090..2665
                     /codon_start=1
                     /product="human Rho family small GTPase Cdc42"
                     /label=Cdc42
                     /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNCAVTVM
                     IGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH
                     CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK
                     GLKNVFDEAILAALEPPEPKKSRRCVLL"
     polyA_signal    2795..2916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2886..2905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2923..3378)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3060..3081)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3272..3291
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3405..3509
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3511..3868
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3511..3702
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3534..3554)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3719..3854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3903..4697
                     /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             3903..4697
                     /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(3957..3976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4567..4586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4885..4904)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4929..4976
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
       61 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      181 gccgtagtta ggccaccact tcaagaactc tgtatcaccg cctacatacc tcgctctgct
      241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
      721 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      781 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      841 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      901 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      961 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1021 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1081 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     1141 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1201 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1261 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
     1321 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1381 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1441 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1501 cgtgccctgg cccaccctcg tgaccaccct gacctggggc gtgcagtgct tcagccgcta
     1561 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1621 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1681 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1741 caacatcctg gggcacaagc tggagtacaa ctacatcagc cacaacgtct atatcaccgc
     1801 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     1861 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1921 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1981 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2041 cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattcca tgcagacaat
     2101 taagtgtgtt gttgtgggcg atggtgccgt tggtaaaaca tgtctcctga tatcctacac
     2161 aacaaacaaa tttccatctg agtatgtacc gactgttttt gacaactgtg cagtcacggt
     2221 tatgattggt ggagagccat atacccttgg actttttgat actgcagggc aagaggatta
     2281 tgacagatta cgaccgctga gttatccaca aacagatgta tttctagtct gtttttcagt
     2341 ggtctctcca tcctcatttg aaaatgtgaa ggaaaagtgg gtgcctgaga taactcacca
     2401 ctgtccaaag actcctttct tgcttgttgg gacccaaatt gatctccgag atgacccctc
     2461 tactatcgag aaacttgcca agaacaaaca gaagcctatc actccagaga ctgctgaaaa
     2521 gctggcccgt gacctgaagg cggtcaagta tgtggagtgt tctgcactca cacagaaagg
     2581 cctaaagaat gtatttgacg aagcaatatt ggctgccctg gagcctccag aaccgaagaa
     2641 gagccgcagg tgtgtgctgc tatgaggatc caccggatct agataactga tcataatcag
     2701 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2761 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2881 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2941 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3001 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3061 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3121 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3181 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3241 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3301 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3361 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3481 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3541 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3601 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3661 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3721 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3841 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3901 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3961 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4021 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4081 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4141 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4201 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4261 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4321 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4381 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4441 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4501 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4561 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4621 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4681 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4741 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4801 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4861 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4921 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4981 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5041 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5101 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5161 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5281 cgt
//
