LOCUS       sequence-389822-        6604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-GGA2 fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6604)
  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 6604)
  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-C2-GGA2
FEATURES             Location/Qualifiers
     source          1..6604
                     /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    3392..3513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3429..3448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3483..3502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3520..3975)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3657..3678)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3869..3888
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4002..4106
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4108..4465
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4108..4299
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4131..4151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4316..4451
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4500..5294
                     /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             4500..5294
                     /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(4554..4573)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5164..5183
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5482..5501)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5526..5573
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5902..6490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6391..6410
                     /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 ccggccggac tcagatctcg agctcaagct tcgaattcat ggcggcgacc
     1381 gcggtggcgg cggctgtggc gggaaccgag tcggcccagg gtcccccggg cccgacagcg
     1441 tcgctggagc tgtggctgaa caaagccaca gacccaagca tgtcggaaca ggattggtca
     1501 gctatccaga atttctgtga gcaggtgaac actgacccca atggccccac acatgcgccc
     1561 tggctgctgg cccacaagat ccagtctccg caagagaagg aagctcttta tgccttaacg
     1621 gtgctggaga tgtgcatgaa ccactgtggg gagaagttcc acagcgaggt ggccaaattt
     1681 cgtttcctga acgaactgat caaagtgttg tccccaaagt acctggggtc ctgggccaca
     1741 ggaaaagtta aaggaagagt cattgaaata ctcttcagtt ggacagtctg gtttccggaa
     1801 gacatcaaga ttcgagacgc ttatcagatg ctgaagaaac aaggaattat aaaacaagac
     1861 cctaaactac cagtggataa aatcttaccc ccaccatctc cctggcccaa gagctccatc
     1921 tttgatgctg atgaagaaaa gtccaagctt ctgacaaggc ttctaaagag caaccacccc
     1981 gaggaccttc aggctgcaaa ccggttaatc aagaatttgg tcaaggagga acaagaaaaa
     2041 tcggagaagg tgtccaagag ggtcagtgcg gtggaggaag tgcgaagcca tgtgaaggtg
     2101 ctgcaggaga tgctgagcat gtaccgcagg ccagggcagg ccccgcccga ccaggaggcc
     2161 ctgcaggtcg tgtatgagag gtgtgaaaag ctgcggccca cgctgttccg gttggcgagt
     2221 gacaccactg atgacgatga tgcactcgcg gaaattctcc aggcaaatga cctcctcacc
     2281 caaggagttc tgctgtacaa acaggtgatg gagggccggg tcacctttgg aaacagagtg
     2341 accagctcat tgggagacat ccctgtctcc agagtctttc agaatccagc aggctgcatg
     2401 aagacctgcc ccctgattga cttggaggtg gacaatggac ctgcgcagat ggggactgtg
     2461 gtgccatctt tgcttcatca ggacctggca gccttgggaa tcagtgatgc tcctgttaca
     2521 ggcatggttt ctggtcagaa ttgctgtgag gaaaagagga atccctcctc cagcacgctg
     2581 ccaggcggtg gtgttcagaa cccttctgca gacaggaatt tgctggacct cctctcacca
     2641 cagccagctc cgtgccctct gaattatgtt tcgcagaaaa gtgtccccaa ggaagtgcca
     2701 ccaggtacta agtcctctcc aggttggtcc tgggaggctg gcccgttggc tccttcccca
     2761 tcttcacaga atacacctct ggctcaagtg tttgtccctt tggagtctgt taagcccagc
     2821 agcctgccgc ctctcattgt gtatgaccgg aatggattca gaattctgct ccacttctcc
     2881 cagacgggag cccctgggca cccagaggta caggtgctgc tcttgaccat gatgagcacg
     2941 gctccccagc ctgtctggga tatcatgttt caagtggctg tgccaaagtc aatgagagtg
     3001 aagctgcagc cggcatccag ctccaagctt cctgcattca gtcctttgat gcctccagct
     3061 gtgatatctc agatgctgct gcttgacaat ccacacaaag aacctatccg cttacggtac
     3121 aagctgacat tcaaccaagg tggacagcct ttcagcgaag taggagaagt gaaagacttc
     3181 ccagacctgg ctgtcttggg cgcagcctaa gcggccgcta aactataagc cgaattctgc
     3241 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca
     3301 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3361 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3421 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3481 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     3541 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3601 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3661 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3721 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3781 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3841 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3901 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3961 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4021 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4081 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4141 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4201 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4261 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4321 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4381 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4441 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4501 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4561 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4621 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4681 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4741 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4801 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4861 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4921 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4981 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5041 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5101 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5161 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5221 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5281 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5341 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5401 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5461 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5521 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5581 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5641 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5701 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5761 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5821 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5881 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5941 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6001 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6061 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6121 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6181 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6241 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6301 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6361 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6421 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6481 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6541 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6601 gcat
//
