LOCUS       sequence-176867-        5425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-tagged human Rab27B N133I in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5425)
  AUTHORS   Hendrix A, Maynard D, Pauwels P, Braems G, Denys H, Van den Broecke 
            R, Lambert J, Van Belle S, Cocquyt V, Gespach C, Bracke M, Seabra 
            MC, Gahl WA, De Wever O, Westbroek W
  TITLE     Effect of the secretory small GTPase Rab27B on breast cancer growth,
            invasion, and metastasis.
  JOURNAL   J Natl Cancer Inst. 2010 Jun 16;102(12):866-80. doi: 
            10.1093/jnci/djq153. Epub 2010 May 18.
  PUBMED    20484105
REFERENCE   2  (bases 1 to 5425)
  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: 
            "10.1093/jnci/djq153"; journalName: "J Natl Cancer Inst"; date: 
            "2010-06-16- 16"; volume: "102"; issue: "12"; pages: "866-80"
COMMENT     Sequence Label: GFP-Rab27B N133I
FEATURES             Location/Qualifiers
     source          1..5425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        771..1074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1075..1278
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1229..1249
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1317..1326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1323..1325,1326..1328,1329..2039)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1323..1325,1326..1328,1329..2039)
                     /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(1368..1389)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1629..1648)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1976..1997
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2923..3044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2960..2979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3014..3033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3051..3506)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3188..3209)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3400..3419
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3533..3637
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3639..3996
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3639..3830
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3662..3682)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3847..3982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4031..4825
                     /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             4031..4825
                     /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(4085..4104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4695..4714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5013..5032)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5057..5104
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
      721 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      781 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      841 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      901 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      961 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1021 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1081 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1141 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     1201 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     1261 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca
     1321 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1381 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1441 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1501 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     1561 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1621 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1681 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1741 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1801 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1861 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1921 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1981 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2041 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc accaagacca
     2101 tcactatgac cgatggagac tatgattatc tgatcaaact cctggccctc ggggattcag
     2161 gggtggggaa gacaacattt ctttatagat acacagataa taaattcaat cccaaattca
     2221 tcactacagt aggaatagac tttcgggaaa aacgtgtggt ttataatgca caaggaccga
     2281 atggatcttc agggaaagca tttaaagtgc atcttcagct ttgggacact gcgggacaag
     2341 agcggttccg gagtctcacc actgcatttt tcagagacgc catgggcttc ttattaatgt
     2401 ttgacctcac cagtcaacag agcttcttaa atgtcagaaa ctggatgagc caactgcaag
     2461 caaatgctta ttgtgaaaat ccagatatag tattaattgg catcaaggca gacctaccag
     2521 atcagaggga agtcaatgaa cggcaagctc gggaactggc tgacaaatat ggcataccat
     2581 attttgaaac aagtgcagca actggacaga atgtggagaa agctgtagaa acccttttgg
     2641 acttaatcat gaagcgaatg gaacagtgtg tggagaagac acaaatccct gatactgtca
     2701 atggtggaaa ttctggaaac ttggatgggg aaaagccacc agagaagaaa tgtatctgct
     2761 agactctaca tagaaactga acatcaagaa cccggatcca ccggatctag ataactgatc
     2821 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2881 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2941 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3001 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     3061 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     3121 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     3181 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     3241 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     3301 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     3361 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     3421 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3481 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3541 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3601 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3661 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3721 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3841 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3901 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3961 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     4021 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4081 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4141 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4201 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4261 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4321 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4381 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4441 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4501 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4561 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4621 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4681 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4741 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4801 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4861 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4921 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4981 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     5041 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     5101 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     5161 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     5221 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     5281 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5341 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5401 tcagaccccg tagaaaagat caaag
//
