LOCUS       sequence-159112-        5358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse Rab21 mutant that has defects in the binding of GTP (dominant 
            negative). Generated from pEGFP-Rab21 by site-directed mutagenesis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5358)
  AUTHORS   Pellinen T, Arjonen A, Vuoriluoto K, Kallio K, Fransen JA, Ivaska J
  TITLE     Small GTPase Rab21 regulates cell adhesion and controls endosomal 
            traffic of beta1-integrins.
  JOURNAL   J Cell Biol. 2006 Jun 5;173(5):767-80.
  PUBMED    16754960
REFERENCE   2  (bases 1 to 5358)
  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 
            Biol."; date: "2006-06-5"; volume: "173(5)"; pages: "767-80"
COMMENT     Sequence Label: pEGFP-Rab21-T31N (DN)
FEATURES             Location/Qualifiers
     source          1..5358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    117..238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(154..173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     208..227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(245..700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(382..403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     594..613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        727..831
                     /gene="bla"
                     /label=AmpR promoter
     promoter        833..1190
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        833..1024
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(856..876)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1041..1176
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1225..2019
                     /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             1225..2019
                     /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(1279..1298)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1889..1908
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2207..2226)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2251..2298
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2627..3215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3116..3135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3390..3693
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3694..3897
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3848..3868
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3936..3945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3942..3944,3945..3947,3948..4658)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3942..3944,3945..3947,3948..4658)
                     /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(3987..4008)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4248..4267)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4595..4616
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctagataact gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
       61 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
      121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
      241 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
      301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
      361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
      421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
      481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
      541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
      601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
      661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
      721 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      781 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
      841 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
      901 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
      961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     1021 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     1081 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     1141 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     1201 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     1261 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     1321 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     1381 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     1441 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     1501 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     1561 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     1621 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     1681 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     1741 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     1801 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     1861 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     1921 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     1981 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     2041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     2101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     2161 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     2221 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     2281 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     2341 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     2401 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     2461 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2521 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2761 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3301 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca
     3361 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3421 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     3481 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     3541 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     3601 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     3661 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     3721 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     3781 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     3841 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     3901 ttagtgaacc gtcagatccg ctagcgctac cggtcgccac catggtgagc aagggcgagg
     3961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4081 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     4141 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4381 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4441 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4561 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4621 ccgccgggat cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc
     4681 aagcttcgat ggctgcggcc ggcggcgggg cggcggcggc ggcgggccga gcctactcct
     4741 tcaaggtggt gctgctcggg gaaggctgcg tggggaagaa ctcgctggtg ctgcgctact
     4801 gcgagaacaa gttcaacgac aagcacatca ccaccctgca ggcatctttc ttaacaaaga
     4861 agttaaatat tggtggaaaa agagtaaacc ttgccatatg ggacacagca ggtcaggaga
     4921 gattccacgc actgggccca atttactacc gagattcgaa cggagctatt ttagtttatg
     4981 atgtaacgga tgaagattcc tttcagaagg taaaaaactg ggtcaaagaa ttacgaaaaa
     5041 tgttgggaaa tgagatctgt ttgtgtatag ttggtaataa aatagacttg gaaaaggaga
     5101 ggcacgtctc tattcaagaa gcagaatcgt atgcagagtc tgtgggagcc aagcattacc
     5161 atacttccgc taaacagaac aaaggcattg aggagctctt tctcgacctt tgtaaaagaa
     5221 tgatagagac agcccaggtg gatgagagag caaaaggcaa tggctcgagt caagccgggg
     5281 ctgcaaggcg aggtgtgcag atcatcgacg acgagcctca agctcagagc agcggagggt
     5341 gctgttcctc cggctaat
//
