LOCUS       sequence-372738-        5082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFP-Rock1 GTPase-binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5082)
  AUTHORS   Budnar S, Husain KB, Gomez GA, Naghibosadat M, Varma A, Verma S, 
            Hamilton NA, Morris RG, Yap AS
  TITLE     Anillin Promotes Cell Contractility by Cyclic Resetting of RhoA 
            Residence Kinetics
  JOURNAL   Dev Cell. 2019 Jun 17;49(6):894-906.e12. doi: 
            10.1016/j.devcel.2019.04.031. Epub 2019 May 16.
  PUBMED    31105010
REFERENCE   2  (bases 1 to 5082)
  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: 
            "10.1016/j.devcel.2019.04.031"; journalName: "Dev Cell"; date: 
            "2019-06-17- 17"; volume: "49"; issue: "6"; pages: "894-906.e12"
COMMENT     Sequence Label: GFP-Rock1-GBD (840-110 aa) in pEGFPN1
FEATURES             Location/Qualifiers
     source          1..5082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        70..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        374..577
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     528..548
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(622..624,625..627,628..1338)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(622..624,625..627,628..1338)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(667..688)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(928..947)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1275..1296
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1879..2000
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1916..1935)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1970..1989
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2007..2462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2144..2165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2356..2375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2489..2593
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2595..2952
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2595..2786
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2618..2638)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2803..2938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2987..3781
                     /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             2987..3781
                     /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(3041..3060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3651..3670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3969..3988)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4013..4060
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4389..4977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4878..4897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
       61 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      121 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      181 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      241 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      301 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      361 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      421 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      481 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      541 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      601 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      661 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      721 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      781 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      841 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      901 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      961 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1021 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1081 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1141 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1201 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1261 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1321 atggacgagc tgtacaagtc cggactcaga tctcgagctc agctggcgcg agggattctg
     1381 gaagaacagt attttgagtt gactcaggaa agcaagaaag ctgcttcaag aaatagacaa
     1441 gagattacag ataaagatca tactgttagt cggcttgaag aaacaaacag tgtgctaacc
     1501 aaagatattg aaatgctaag aaaagaaaat gaagagctaa atgaaagaat gaggacagca
     1561 gaagaagaat ataaattgaa gaaggaggaa gaaataaata atcttaaggc tgcttttgaa
     1621 aagaatatca gtaccgaacg gacccttaaa acacaggctg ttaacaagtt ggcagaaata
     1681 atgaatcgca aagattttaa aattgatcga aagaaagcta atacacaaga tttaagaaaa
     1741 aaagaatagg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     1801 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1861 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1921 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1981 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2041 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2101 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2161 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2221 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2281 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2341 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2401 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2461 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2581 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2641 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2701 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2761 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2821 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2881 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2941 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3001 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3061 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3121 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3181 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3241 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3301 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3361 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3421 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3481 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3541 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3601 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3661 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3721 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3781 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3841 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3901 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3961 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4021 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4081 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4141 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4201 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4261 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4321 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4381 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4441 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4501 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4561 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4621 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4681 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4741 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4801 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4861 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4921 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4981 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5041 tttcctgcgt tatcccctga ttctgtggat aaccgtatta cc
//
