LOCUS       sequence-176862-        5425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-tagged human Rab27B 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
FEATURES             Location/Qualifiers
     source          1..5425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      16..604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     505..524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        779..1082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1083..1286
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1237..1257
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1325..1334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1331..1333,1334..1336,1337..2047)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1331..1333,1334..1336,1337..2047)
                     /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(1376..1397)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1637..1656)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1984..2005
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2931..3052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2968..2987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3022..3041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3059..3514)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3196..3217)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3408..3427
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3541..3645
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3647..4004
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3647..3838
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3670..3690)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3855..3990
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4039..4833
                     /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             4039..4833
                     /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(4093..4112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4703..4722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5021..5040)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5065..5112
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
       61 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
      121 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
      181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
      721 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      781 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      841 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      901 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      961 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1021 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1081 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1141 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1201 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1261 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
     1321 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1381 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1441 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1501 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1561 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1621 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1681 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1741 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1801 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1861 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1921 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1981 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2041 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccac
     2101 caagaccatc actatgaccg atggagacta tgattatctg atcaaactcc tggccctcgg
     2161 ggattcaggg gtggggaaga caacatttct ttatagatac acagataata aattcaatcc
     2221 caaattcatc actacagtag gaatagactt tcgggaaaaa cgtgtggttt ataatgcaca
     2281 aggaccgaat ggatcttcag ggaaagcatt taaagtgcat cttcagcttt gggacactgc
     2341 gggacaagag cggttccgga gtctcaccac tgcatttttc agagacgcca tgggcttctt
     2401 attaatgttt gacctcacca gtcaacagag cttcttaaat gtcagaaact ggatgagcca
     2461 actgcaagca aatgcttatt gtgaaaatcc agatatagta ttaattggca acaaggcaga
     2521 cctaccagat cagagggaag tcaatgaacg gcaagctcgg gaactggctg acaaatatgg
     2581 cataccatat tttgaaacaa gtgcagcaac tggacagaat gtggagaaag ctgtagaaac
     2641 ccttttggac ttaatcatga agcgaatgga acagtgtgtg gagaagacac aaatccctga
     2701 tactgtcaat ggtggaaatt ctggaaactt ggatggggaa aagccaccag agaagaaatg
     2761 tatctgctag actctacata gaaactgaac atcaagaacc cggatccacc ggatctagat
     2821 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2881 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     3061 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3121 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     3181 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     3241 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     3301 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     3361 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     3421 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3481 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3541 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3601 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3661 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3721 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3841 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3901 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3961 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4021 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4081 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4141 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4201 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     4261 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4321 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4381 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4441 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4501 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4561 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4621 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4681 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4741 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4801 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4861 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4921 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4981 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5041 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5101 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     5161 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     5221 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     5281 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5341 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5401 actgagcgtc agaccccgta gaaaa
//
