LOCUS       sequence-13783-e        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  AUTHORS   Murakoshi H, Wang H, Yasuda R
  TITLE     Local, persistent activation of Rho GTPases during plasticity of 
            single dendritic spines.
  JOURNAL   Nature. 2011 Apr 7;472(7341):100-4. doi: 10.1038/nature09823. Epub 
            2011 Mar 20.
  PUBMED    21423166
REFERENCE   2  (bases 1 to 5268)
  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: 
            "10.1038/nature09823"; journalName: "Nature"; date: "2011-04-7- 7"; 
            volume: "472"; issue: "7341"; pages: "100-4"
COMMENT     Sequence Label: mEGFP-Cdc42-C1
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             281..856
                     /codon_start=1
                     /product="human Rho family small GTPase Cdc42"
                     /label=Cdc42
                     /translation="MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVM
                     IGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHH
                     CPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQK
                     GLKNVFDEAILAALEPPEPKKSRRCVLL"
     polyA_signal    986..1107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1023..1042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1077..1096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1114..1569)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1251..1272)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1463..1482
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1596..1700
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1702..2059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1702..1893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1725..1745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1910..2045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2094..2888
                     /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             2094..2888
                     /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(2148..2167)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2758..2777
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3076..3095)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3120..3167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3496..4084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3985..4004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4259..4562
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4563..4766
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4717..4737
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4805..4814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4811..4813,4814..4816,4817..5268,1..259)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4856..4877)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5117..5136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
       61 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      121 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
      181 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
      241 catggacgag ctgtacaagt ccggactcag atctcgaggg atgcagacaa ttaagtgtgt
      301 tgttgtgggc gatggtgctg ttggtaaaac atgtctcctg atatcctaca caacaaacaa
      361 atttccatcg gaatatgtac cgactgtttt tgacaactat gcagtcacag ttatgattgg
      421 tggagaacca tatactcttg gactttttga tactgcaggg caagaggatt atgacagatt
      481 acgaccgctg agttatccac aaacagatgt atttctagtc tgtttttcag tggtctctcc
      541 atcttcattt gaaaacgtga aagaaaagtg ggtgcctgag ataactcacc actgtccaaa
      601 gactcctttc ttgcttgttg ggactcaaat tgatctcaga gatgacccct ctactattga
      661 gaaacttgcc aagaacaaac agaagcctat cactccagag actgctgaaa agctggcccg
      721 tgacctgaag gctgtcaagt atgtggagtg ttctgcactt acacagaaag gcctaaagaa
      781 tgtatttgac gaagcaatat tggctgccct ggagcctcca gaaccgaaga agagccgcag
      841 gtgtgtgctg ctatgaggat ccaccggatc tagataactg atcataatca gccataccac
      901 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      961 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1081 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     1141 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     1201 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     1261 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     1321 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     1381 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     1441 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     1501 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     1561 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1621 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1681 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     1741 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1801 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     1861 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     1921 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     1981 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2041 aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg
     2101 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2161 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2221 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg
     2281 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2341 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2401 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2461 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2521 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2581 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg
     2641 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2701 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     2761 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     2821 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     2881 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     2941 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3001 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3061 tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3121 ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt
     3181 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     3241 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     3301 agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat
     3361 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3421 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3481 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3541 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3601 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3661 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3721 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3781 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3841 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3901 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3961 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4021 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4081 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4141 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
     4201 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     4261 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     4321 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     4381 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     4441 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     4501 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     4561 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     4621 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     4681 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     4741 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     4801 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4861 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4921 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4981 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     5041 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     5101 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     5161 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5221 cctggggcac aagctggagt acaactacaa cagccacaac gtctatat
//
