LOCUS       sequence-176864-        5425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GFP-tagged human Rab27B Q78L 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 Q78L
FEATURES             Location/Qualifiers
     source          1..5425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        783..1086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1087..1290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1241..1261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1329..1338
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1335..1337,1338..1340,1341..2051)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1335..1337,1338..1340,1341..2051)
                     /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(1380..1401)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1641..1660)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1988..2009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2935..3056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2972..2991)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3026..3045
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3063..3518)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3200..3221)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3412..3431
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3545..3649
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3651..4008
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3651..3842
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3674..3694)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3859..3994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4043..4837
                     /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             4043..4837
                     /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(4097..4116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4707..4726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5025..5044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5069..5116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
      721 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      781 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      841 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      901 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      961 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1021 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1081 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1141 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     1201 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     1261 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
     1321 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1381 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1441 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1501 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1561 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1621 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1681 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1741 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1801 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1861 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1921 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1981 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2041 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     2101 ccaccaagac catcactatg accgatggag actatgatta tctgatcaaa ctcctggccc
     2161 tcggggattc aggggtgggg aagacaacat ttctttatag atacacagat aataaattca
     2221 atcccaaatt catcactaca gtaggaatag actttcggga aaaacgtgtg gtttataatg
     2281 cacaaggacc gaatggatct tcagggaaag catttaaagt gcatcttcag ctttgggaca
     2341 ctgcgggact agagcggttc cggagtctca ccactgcatt tttcagagac gccatgggct
     2401 tcttattaat gtttgacctc accagtcaac agagcttctt aaatgtcaga aactggatga
     2461 gccaactgca agcaaatgct tattgtgaaa atccagatat agtattaatt ggcaacaagg
     2521 cagacctacc agatcagagg gaagtcaatg aacggcaagc tcgggaactg gctgacaaat
     2581 atggcatacc atattttgaa acaagtgcag caactggaca gaatgtggag aaagctgtag
     2641 aaaccctttt ggacttaatc atgaagcgaa tggaacagtg tgtggagaag acacaaatcc
     2701 ctgatactgt caatggtgga aattctggaa acttggatgg ggaaaagcca ccagagaaga
     2761 aatgtatctg ctagactcta catagaaact gaacatcaag aacccggatc caccggatct
     2821 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2881 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2941 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3001 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     3061 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3121 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3181 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3241 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3301 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3361 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3421 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3481 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3661 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4021 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4081 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4141 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4201 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4261 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4321 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4381 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4441 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4501 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4561 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4621 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4681 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4741 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4801 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4861 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4921 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4981 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5041 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5101 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5161 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5221 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5281 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5341 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5401 ttccactgag cgtcagaccc cgtag
//
