LOCUS       sequence-346373-        5680 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively active, nuclear localized RhoG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5680)
  AUTHORS   Mahlandt EK, Kreider-Letterman G, Chertkova AO, Garcia-Mata R, 
            Goedhart J
  TITLE     Cell-based optimization and characterization of genetically encoded 
            location-based biosensors for Cdc42 or Rac activity.
  JOURNAL   J Cell Sci. 2023 May 15;136(10):jcs260802. doi: 10.1242/jcs.260802. 
            Epub 2023 May 25.
  PUBMED    37226883
REFERENCE   2  (bases 1 to 5680)
  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"; journalName: "J Cell 
            Sci."; date: "2023-05-15"; volume: "136(10):jcs260802. doi"; pages: 
            " 10.1242/jcs.260802. Epub 2023 May 25"
COMMENT     Sequence Label: H2A-mTurquoise-RHOG-QL-deltaCAAX
FEATURES             Location/Qualifiers
     source          1..5680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      413..422
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(419..421,422..424,425..1135)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(464..485)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(725..744)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1072..1093
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1839..1960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1876..1895)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1930..1949
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1967..2422)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2104..2125)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2316..2335
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2449..2553
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2555..2912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2555..2746
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2578..2598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2763..2898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2947..3741
                     /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             2947..3741
                     /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(3001..3020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3611..3630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3929..3948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3973..4020
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4349..4937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4838..4857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5112..5415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5416..5619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5570..5590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aattctgatg tcgggacgcg gcaagcaggg aggcaaagct cgcgccaaag ccaagacccg
       61 ctcttctcgt gccggtctcc agttccccgt gggccgagtg caccgactgc tccgcaaggg
      121 caactatgct gagcgggtcg gggccggcgc gccggtgtac ctggcggcgg tgctggagta
      181 cctgactgcc gagatcctgg agctggcggg caacgccgcc cgcgacaaca agaagacccg
      241 cattatcccg cgccacttgc agctggccat ccgcaacgac gaggagctca acaagctgct
      301 gggcaaagta accatcgctc agggtggtgt cctgcccaac atccaggctg tgctactgcc
      361 caagaagacc gagagtcacc acaaggccaa aggcaaacgg gatccaccgg tcgccaccat
      421 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
      481 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
      541 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
      601 cgtgaccacc ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca
      661 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
      721 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
      781 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
      841 gctggagtac aactacatca gcgacaacgt ctatatcacc gccgacaagc agaagaacgg
      901 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
      961 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1021 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1081 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     1141 actcagatct atgcagagca tcaagtgcgt ggtggtgggt gatggggctg tgggcaagac
     1201 gtgcctgctc atctgctaca caactaacgc tttccccaaa gagtacatcc ccaccgtgtt
     1261 cgacaattac agcgcgcaga gcgcagttga cgggcgcaca gtgaacctga acctgtggga
     1321 cactgcgggc ctggaggagt atgaccgcct ccgtacactc tcctaccctc agaccaacgt
     1381 tttcgtcatc tgtttctcca ttgccagtcc gccgtcctat gagaacgtgc ggcacaagtg
     1441 gcatccagag gtgtgccacc actgccctga tgtgcccatc ctgctggtgg gcaccaagaa
     1501 ggacctgaga gcccagcctg acaccctacg gcgcctcaag gagcagggcc aggcgcccat
     1561 cacaccgcag cagggccagg cactggccaa gcagatccac gctgtgcgct acctcgaatg
     1621 ctcagccctg caacaggatg gtgtcaagga agtgttcgcc gaggctgtcc gggctgtgct
     1681 caaccccacg ccgatcaagc gtgggcggtc cagctaagcg gccgcgactc tagatcataa
     1741 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1801 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1861 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1921 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     1981 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2041 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2101 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2161 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2221 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2281 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2341 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2401 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2521 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2821 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2881 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2941 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3001 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3061 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3121 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3181 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3241 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3301 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3361 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3421 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3481 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3541 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3601 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3661 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3721 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3781 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3841 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3901 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3961 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4021 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4081 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4141 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4201 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4261 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4321 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4381 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4441 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4501 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4561 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4621 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4681 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4741 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4801 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4861 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4921 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4981 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5041 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     5101 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5161 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5221 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5281 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     5341 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     5401 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     5461 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     5521 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     5581 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     5641 cgctagcgct accggactca gatctcgagc tcaagcttcg
//
