LOCUS       sequence-195050-        8874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Rock2-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8874)
  AUTHORS   Priya R, Gomez GA, Budnar S, Verma S, Cox HL, Hamilton NA, Yap AS
  TITLE     Feedback regulation through myosin II confers robustness on RhoA 
            signalling at E-cadherin junctions.
  JOURNAL   Nat Cell Biol. 2015 Oct;17(10):1282-93. doi: 10.1038/ncb3239. Epub 
            2015 Sep 14.
  PUBMED    26368311
REFERENCE   2  (bases 1 to 8874)
  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/ncb3239"; 
            journalName: "Nat Cell Biol"; date: "2015-10"; volume: "17"; issue: 
            "10"; pages: "1282-93"
COMMENT     Sequence Label: GFP-mROCK2
FEATURES             Location/Qualifiers
     source          1..8874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      136..724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     625..644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        899..1202
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1203..1406
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1357..1377
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1445..1454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1451..1453,1454..1456,1457..2167)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1451..1453,1454..1456,1457..2167)
                     /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(1496..1517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1757..1776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2104..2125
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6500..6621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6537..6556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6591..6610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6628..7083)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6765..6786)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6977..6996
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7110..7214
                     /gene="bla"
                     /label=AmpR promoter
     promoter        7216..7573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7216..7407
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7239..7259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7424..7559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7608..8402
                     /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             7608..8402
                     /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(7662..7681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8272..8291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8590..8609)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8634..8681
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
       61 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
      121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
      181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
      841 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      901 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      961 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     1021 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     1081 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1141 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1201 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1261 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1321 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1381 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     1441 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1501 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1561 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1621 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1681 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1741 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1801 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1861 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1921 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1981 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2041 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2101 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2161 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctatgagcc ggcccccgcc
     2221 gacggggaaa atgcccggcg cccccgagac cgcgccgggg gacggggcgg gcgccggccg
     2281 tcagaggaag ctggaggcgc tcatccgaga ccctcgctcg cccatcaacg tggagagtct
     2341 gctggatggc ttaaattcct tggtccttga tttggatttt cctgctttga ggaaaaataa
     2401 aaatatagat aacttcttaa atagatatga gaaaattgtg aaaaaaatca gaggtttaca
     2461 gatgaaagcg gaagactatg atgttgtaaa agttattgga agaggtgctt ttggtgaagt
     2521 tcagttggtt cgtcataagg catcacagaa ggtttatgca atgaagcttc ttagtaaatt
     2581 tgaaatgata aaaagatcag attctgcttt tttctgggaa gaaagagata ttatggcctt
     2641 tgccaacagt ccctgggtag ttcagctctt ctgtgccttt caagatgaca ggtacctgta
     2701 catggtgatg gagtacatgc caggcggaga cctcgtcaac cttatgagta attatgatgt
     2761 acctgaaaag tgggccaagt tttacactgc tgaagtagtg cttgcactgg atgcaataca
     2821 ctccatgggc ttaatacaca gagatgtgaa gcctgataac atgctattgg ataaacatgg
     2881 acatctaaag ttagcagatt ttggcacatg tatgaaaatg gatgaaacag gcatggtgca
     2941 ttgcgacaca gcagttggaa ctcccgacta catatcgccc gaggttctga aatcacaagg
     3001 aggtgatggt tactatgggc gagaatgtga ttggtggtct gtaggtgttt tcctttttga
     3061 gatgttagta ggggacactc cattttacgc agattcactt gtaggaacct acagcaaaat
     3121 tatggatcat aaaaattcac tgtgttttcc tgaagataca gaaatttcta aacatgcgaa
     3181 gaatctcata tgtgccttct taacagacag ggaggtacga cttggaagaa atggggtaga
     3241 agaaatcaaa cagcatcctt tctttaagaa tgatcaatgg aattgggata acataagaga
     3301 gacggcagct ccagtggtac ctgagctcag cagtgacata gacagcagca actttgacga
     3361 cattgaggat gacaaaggcg atgtagagac cttcccgatt cctaaagctt ttgtgggaaa
     3421 tcagctgcct tttataggat ttacctactt tagagaaaat ttgttattaa gtgactctcc
     3481 accttgtaga gaaaatgatg caatacaaac aaggaaaagt gaagaaagtc aggagattca
     3541 gaaaaaatta tatgcactag aagaacacct tagcagtgag gtgcaagcca aggaggagct
     3601 ggaacagaag tgcaaatcta ttaatactcg cctagagaaa acagcaaagg agctagaaga
     3661 agagattacc ttacggaaaa gtgtggaatc aacattaaga cagctggaaa gagaaaaggc
     3721 ccttcttcag cacaaaaatg cagagtatca gcggaaagct gatcatgaag ccgacaagaa
     3781 gcggaacttg gaaaatgatg ttaacagctt aaaagatcaa cttgaagatc tgaagaaaag
     3841 gaaccagagc tctcagatat ccactgagaa ggtcaatcag ctccagaaac aactggatga
     3901 agctaatgct ttgctgagaa cagagtctga tactgcagca cggttaagaa aaacccaggc
     3961 agaaagttcc aaacagattc agcagctgga atctaacaat agagatctac aagataaaaa
     4021 ctgcctgctt gagactgcca agttaaaact tgaaaaggaa tttatcaatc ttcagtcagc
     4081 tttagaatct gaaagaaggg accggaccca tggatcagag ataattaatg atttacaagg
     4141 tagaatatct gggttggaag aagatttgaa gactggcaaa gccttactag caaaagtaga
     4201 gctggagaag agacagctgc aggagaagct cactgactta gagaaggaaa aaagcaacat
     4261 ggaaatagat atgacatatc aactgaaagt tatacagcag agtttagaac aagaagaagc
     4321 tgaacacaag accacaaaag cacggctagc ggataaaaat aagatctatg aatccattga
     4381 agaagctaaa tcagaagcca tgaaagaaat ggagaagaag ctgttggaag agaggtcttt
     4441 aaagcagaaa gtggagaacc tgctcctgga agccgagaaa agatgctcta tattggactg
     4501 tgacctcaaa cagtctcagc agaagctaaa tgagctgctg aagcagaaag atgtgctgaa
     4561 cgaggatgtt agaaatttga cattaaaaat agagcaggaa actcagaagc gttgtcttat
     4621 gcaaaatgac ctgaagatgc agacccaaca agttaacacc ctaaaaatgt cagaaaagca
     4681 aataaaacaa gaaaataatc atcttatgga aatgaaaatg aacttggaaa agcaaaatac
     4741 tgaacttcga aaagagcggc aggatgcaga tgggcagatg aaggaactgc aagaccaact
     4801 tgaagcagag cagtatttct caaccctcta taaaacacaa gttagagaac ttaaagaaga
     4861 aaatgaagag aagacaaaac tttgtaaaga gttgcaacag aagaagcagg atttacagga
     4921 tgaaagagac tctttggctg cccagctgga aattaccctt accaaagcag attctgagca
     4981 gctggctcgt tccattgctg aagaacaata ctctgatttg gaaaaagaga agatcatgaa
     5041 agagctggag atcaaagaga tgatggctag acacaaacaa gaacttactg aaaaggatac
     5101 tacaattgca tctcttgaag aaacaaatag gacactaact agcgatgttg caaatcttgc
     5161 aaatgagaaa gaagaattaa acaacaagct gaaagatagc caagaacaac tgtccaagtt
     5221 gaaagatgaa gagatgagtg cagcagctat taaagcacag tttgagaagc agcttctgaa
     5281 tgagagaact ctcaagactc aagctgtgaa taagttggcg gagatcatga atcgaaaaga
     5341 acctgtcaag cgtggtagtg acacagatgt gcgaagaaaa gagaaggaaa acagaaagct
     5401 acatatggag cttaagtctg aacgtgaaaa attgacacaa cagatgatca aatatcagaa
     5461 agaactgaat gaaatgcagg ctcaaatagc tgaagagagc caaattcgaa tagaactcca
     5521 gatgaccctg gacagtaaag acagcgacat tgaacagctg cggtcacagc tccaggcctt
     5581 gcatattggt atggacagtt ccagtatagg cagtggacca ggcgatgctg agcctgatga
     5641 tggatttcca gaatcaagat tagaaggatg gttgtcattg cctgtgcgca acaacactaa
     5701 aaagtttgga tgggttaaaa agtatgtgat tgtaagcagc aagaagattc ttttttatga
     5761 cagcgaacaa gataaagagc agtccaaccc ttacatggtg ttagacatag acaagctatt
     5821 tcacgtccga cctgttaccc agacagatgt atacagagca gatgctaaag aaattccaag
     5881 gatatttcag attctatatg ccaatgaagg agaaagtaag aaggaaccag agtttccagt
     5941 ggaaccagtg ggagaaaaat ctaattatat ttgtcacaaa ggacatgagt ttattcctac
     6001 tctttatcac ttcccaacca actgtgaggc atgtatgaag ccgctgtggc acatgtttaa
     6061 gcctcctcct gctctagagt gccgtaggtg ccacattaaa tgtcataaag atcacatgga
     6121 caagaaggaa gagattatag cgccctgcaa agtttattat gatatatcgt cggcaaagaa
     6181 tctgttgtta ttagcaaatt caacagaaga acagcagaaa tgggttagtc ggttggtgaa
     6241 aaagatacct aaaaagcctc cagctccaga cccttttgcc cgatcatccc ctagaacttc
     6301 aatgaaaata caacaaaacc agtccattcg gcggcccagt cgacagcttg ctccaaacaa
     6361 gccaagctaa ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
     6421 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     6481 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     6541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6601 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     6661 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     6721 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     6781 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     6841 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     6901 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     6961 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7021 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7081 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7201 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     7261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7321 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7381 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     7441 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7501 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7561 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     7621 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     7681 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     7741 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     7801 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     7861 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     7921 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     7981 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     8041 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     8101 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     8161 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     8221 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     8281 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     8341 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     8401 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     8461 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     8521 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     8581 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     8641 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     8701 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     8761 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     8821 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agat
//
