LOCUS       sequence-151386-        6106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruitable mCherry tagged RhoA GEF LARG (DH domain). Expresses in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6106)
  AUTHORS   Wagner E, Glotzer M
  TITLE     Local RhoA activation induces cytokinetic furrows independent of 
            spindle position and cell cycle stage.
  JOURNAL   J Cell Biol. 2016 Jun 20;213(6):641-9. doi: 10.1083/jcb.201603025. 
            Epub 2016 Jun 13.
  PUBMED    27298323
REFERENCE   2  (bases 1 to 6106)
  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.201603025"; journalName: "J Cell Biol"; date: 
            "2016-06-20- 20"; volume: "213"; issue: "6"; pages: "641-9"
COMMENT     Sequence Label: PR_GEF  (2XPDZ-mCherry-Larg (DH)
FEATURES             Location/Qualifiers
     source          1..6106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1246..1298
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1306..1315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1312..2019
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1457..1475)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1725..1744
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2894..3015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2985..3004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3022..3477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3159..3180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3371..3390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3504..3608
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3610..3967
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3610..3801
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3633..3653)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3818..3953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4002..4796
                     /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             4002..4796
                     /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(4056..4075)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4666..4685
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4984..5003)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5028..5075
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5404..5992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5893..5912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatc gataccgtcg acatggcaaa acaagagatt
      661 cgagtgaggg ttgaaaagga cccagaactt ggatttagca tatcaggtgg tgtcgggggt
      721 agaggaaacc cattcagacc tgatgatgat ggtatatttg taacaagggt acaacctgaa
      781 ggaccagcat caaaattact gcagccaggt gataaaatta ttcaggctaa tggctacagt
      841 tttataaata ttgaacatgg acaagcagtg tccttgctaa aaactttcca gaatacagtt
      901 gaactcatca ttgtacgaga agttagtgct ggtggtagtg caggagggag tgctggcggg
      961 gcaaaacaag agattcgagt gagggttgaa aaggacccag aacttggatt tagcatatca
     1021 ggtggtgtcg ggggtagagg aaacccattc agacctgatg atgatggtat atttgtaaca
     1081 agggtacaac ctgaaggacc agcatcaaaa ttactgcagc caggtgataa aattattcag
     1141 gctaatggct acagttttat aaatattgaa catggacaag cagtgtcctt gctaaaaact
     1201 ttccagaata cagttgaact catcattgta cgagaagttt ctagactcga gctcaagctt
     1261 cgaattctgc aatcgacggt accgcgggcc cgggatccac cggtcgccac catggtgagc
     1321 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1381 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1441 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     1501 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     1561 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1621 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     1681 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     1741 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     1801 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     1861 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     1921 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     1981 ggccgccact ccaccggcgg catggacgag ctgtacaagc tcgagctcaa gcttagtgct
     2041 ggtggtagtg caggaggctc ggccggtgga agtgcaggag ggagtgctgg cggggtcgac
     2101 gacaccccac ccaactggca gcagcttgtt agtcgagaag tgttactggg actaaaacct
     2161 tgtgaaatca aaagacagga agtgattaat gaattgttct acactgaaag agctcatgtt
     2221 cgaacactga aggttcttga tcaagtgttc tatcagcgag tatccagaga aggaattctg
     2281 tcaccctcag agctacggaa aattttttca aacttggaag atattcttca acttcatatt
     2341 ggattgaatg aacaaatgaa ggctgttcga aagagaaatg agacctctgt tatcgatcag
     2401 attggggaag atttgctgac atggttcagc ggaccaggag aggagaaatt gaaacatgct
     2461 gctgctacct tttgcagtaa ccaacctttc gccctggaaa tgatcaaatc tcgtcagaaa
     2521 aaggattctc gatttcagac ttttgtgcaa gatgctgaaa gtaatccact gtgtcgtcgt
     2581 cttcaactga aggatattat tcccactcaa atgcaaaggc ttactaagta cccacttctg
     2641 ttggataata ttgccaaata cacagaatgg ccaacagaaa gggagaaggt gaagaaagct
     2701 gcagatcact gtcgtcagat cttaaattat gtaaatcagg ctgtcaagga ggcagaaaac
     2761 aagcagcgct aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     2821 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2881 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3001 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3061 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3121 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3181 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3241 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3301 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3361 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3421 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3601 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3841 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3901 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3961 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4021 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4081 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4141 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4201 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4261 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4321 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4381 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4441 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4501 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4561 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4621 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4681 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4741 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4801 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4861 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4921 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4981 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5041 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5101 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5161 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5221 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5281 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5341 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5401 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5461 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5521 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5581 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5641 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5701 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5761 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5821 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5881 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5941 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6001 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6061 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
