LOCUS       sequence-389842-        7452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-Rabaptin-5 fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7452)
  AUTHORS   Mattera R, Arighi CN, Lodge R, Zerial M, Bonifacino JS
  TITLE     Divalent interaction of the GGAs with the Rabaptin-5-Rabex-5 
            complex.
  JOURNAL   EMBO J. 2003 Jan 2;22(1):78-88. doi: 10.1093/emboj/cdg015.
  PUBMED    12505986
REFERENCE   2  (bases 1 to 7452)
  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"; journalName: 
            "EMBO J"; date: "2003-01-2- 2"; volume: "22"; issue: "1"; pages: 
            "78-88"
COMMENT     Sequence Label: pEGFP-C3-Rabaptin-5
FEATURES             Location/Qualifiers
     source          1..7452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4240..4361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4277..4296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4331..4350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4368..4823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4505..4526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4717..4736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4850..4954
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4956..5313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4956..5147
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4979..4999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5164..5299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5348..6142
                     /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             5348..6142
                     /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(5402..5421)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6012..6031
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6330..6349)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6374..6421
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6750..7338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7239..7258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     1381 gccccggctt cccagcctga cgtttctctt cagcaacggg tagcagaatt ggaaaaaatt
     1441 aatgcagaat ttttacgtgc acaacagcag cttgaacaag aatttaatca aaagagagca
     1501 aaatttaagg agttatattt ggctaaagag gaggatctga agaggcaaaa tgcagtatta
     1561 caagctgcac aagatgattt gggacacctt cgaacccagc tgtgggaagc tcaagcagag
     1621 atggagaata ttaaggcgat tgccacagtc tctgagaaca ccaagcaaga agctatagat
     1681 gaagtgaaaa gacagtggag agaagaagtt gcttcacttc aggctgttat gaaagaaaca
     1741 gttcgtgact atgagcacca gttccacctt aggctggagc aggagcgaac acagtgggca
     1801 cagtatagag aatccgcaga gagggaaata gctgatttaa gaagaaggct gtctgaaggt
     1861 caagaggagg aaaatttaga aaatgaaatg aaaaaggccc aagaggatgc tgagaaactt
     1921 cggtccgttg tgatgccaat ggaaaaggaa attgcagctt tgaaggataa actgacagag
     1981 gctgaagaca aaattaaaga gctggaggcc tcaaaggtta aagaactgaa tcattatctg
     2041 gaagctgaga aatcttgtag gactgatcta gagatgtatg tagctgtttt gaatactcag
     2101 aaatctgttc tacaggaaga tgctgagaaa ctgcggaaag aattgcatga agtttgccat
     2161 ctcttggagc aagagcgaca acaacacaac cagttaaaac atacgtggca gaaggccaat
     2221 gaccagtttc tggaatctca gcgtttactg atgagagaca tgcagcgaat ggagattgtg
     2281 ctaacttcag aacagctccg acaagttgaa gaactgaaga agaaagatca ggaggatgat
     2341 gaacaacaaa gactcaataa gagaaaggat cacaaaaaag cagatgttga ggaagaaata
     2401 aaaataccag tagtgtgtgc tttaactcaa gaagaatctt cagcccagtt atcaaatgaa
     2461 gaggagcatt tagacagcac ccgtggctca gttcattcct tagatgcagg cttgctgttg
     2521 ccatctggag atcctttcag taaatcggac aatgacatgt ttaaagatgg actcaggaga
     2581 gcacagtcta cagacagctt gggaacctcg ggctcattgc aatccaaagc tttaggctat
     2641 aactacaaag caaaatctgc tggaaacctg gacgagtcag attttggacc actggtagga
     2701 gcagattcag tgtctgagaa ctttgatact gcatcccttg ggtcactcca gatgccaagt
     2761 gggtttatgt taaccaaaga tcaggaaaga gcaatcaagg cgatgacacc agaacaagaa
     2821 gagacagcgt ccctcctctc cagcgttacc cagggcatgg agagtaccta tgtgtcccct
     2881 agtggttatc gtttagttag tgaaacagaa tggaatctct tgcagaaaga ggtacataat
     2941 gctggaaata aacttggtag acgttgtgat atgtgttcca attacgaaaa acagttacaa
     3001 ggaattcaga ttcaggaggc tgaaacgaga gaccaggtga aaaaactaca gctgatgcta
     3061 aggcaagcta atgaccagtt agagaagaca atgaaagata agcaggagct ggaagacttc
     3121 ataaagcaaa gcagcgaaga ttcgagtcac cagatctctg cactcgtcct aagagcccag
     3181 gcctccgaga tcttacttga aaagttacag caggggcttt cccaggcaaa gagggatgtt
     3241 caggaacaga tggcggtgct gatgcagtca cgggaacagg tttcagaaga gctggtgagg
     3301 ttacagaaag ataatgacag tctccaggga aagcacagcc tgcatgtgtc attacagcaa
     3361 gcagaagact tcatcctccc agacactaca gaggcactgc gggagttggt attaaaatac
     3421 cgtgaggaca tcattaatgt gcggacagca gcagaccacg tagaagaaaa gctgaaggct
     3481 gagatacttt tcctaaaaga gcagatccaa gcagaacagt gtttaaaaga aaatcttgaa
     3541 gaaactctgc aactagaaat agaaaactgc aaggaggaaa tagcttctat ttctagccta
     3601 aaagctgaat tagaaagaat aaaagtggaa aaaggacagt tggagtccac attaagagag
     3661 aagtctcaac agcttgagag tcttcaggaa ataaagatca gtttggaaga gcagttaaag
     3721 aaagagactg ctgctaaggc taccgttgaa cagctaatgt ttgaagagaa gaacaaagct
     3781 cagagattac agacagaatt agatgtcagt gagcaagtcc agagagattt tgtaaagctt
     3841 tcacagaccc ttcaggtgca gttagagcgg atccggcaag ctgactcctt ggagagaatc
     3901 cgggcaattc tgaatgatac taaactgaca gacattaacc agcttcctga gacatgacac
     3961 cctcatggca ggattctagc ctgcactttg ggtttttaac tcatctttag agcaacagta
     4021 attattattt aactcttaac tgaagaaaga gaagtcacaa caaaaggaag actggagaaa
     4081 tgcttacttc tagagtcgac cgggcggccg ggatccaccg gatctagata actgatcata
     4141 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4201 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4261 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4321 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     4381 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     4441 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     4501 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     4561 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     4621 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     4681 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     4741 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4801 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     4981 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5041 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5101 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5161 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5221 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     5281 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     5341 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5401 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5461 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5521 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5581 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5641 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5701 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5761 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5821 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     5881 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5941 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6001 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6061 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6121 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6181 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6241 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6301 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     6361 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     6421 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     6481 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     6541 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     6601 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6661 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6721 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6781 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6841 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6901 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6961 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7021 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7081 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7141 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7201 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7261 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7321 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7381 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7441 accgccatgc at
//
