LOCUS       sequence-350945-        6639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GGA1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6639)
  AUTHORS   Puertollano R, van der Wel NN, Greene LE, Eisenberg E, Peters PJ, 
            Bonifacino JS
  TITLE     Morphology and dynamics of clathrin/GGA1-coated carriers budding 
            from the trans-Golgi network.
  JOURNAL   Mol Biol Cell. 2003 Apr;14(4):1545-57. doi: 10.1091/mbc.02-07-0109.
  PUBMED    12686608
REFERENCE   2  (bases 1 to 6639)
  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: "10.1091/mbc"; 
            journalName: "Mol Biol Cell"; date: "2003-04"; volume: "14"; issue: 
            "4"; pages: "1545-57"
COMMENT     Sequence Label: GFP-GGA1
FEATURES             Location/Qualifiers
     source          1..6639
                     /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    3427..3548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3464..3483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3518..3537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3555..4010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3692..3713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3904..3923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4037..4141
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4143..4500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4143..4334
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4166..4186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4351..4486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4535..5329
                     /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             4535..5329
                     /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(4589..4608)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5199..5218
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5517..5536)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5561..5608
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5937..6525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6426..6445
                     /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 ggagccggag
     1381 actctggagg cgcgaatcaa tagagccacg aaccccctga acaaggagct cgactgggcc
     1441 agcatcaacg gcttctgcga gcagctcaac gaggactttg aggggcctcc actcgccacc
     1501 cggctgctgg cccacaagat ccagtcccca caggagtggg aggcgatcca ggccttgacg
     1561 gtgctggaaa catgcatgaa gagctgcggc aagcggttcc acgacgaagt gggcaagttc
     1621 cgctttctca acgagctcat caaggtcgtg tctcccaagt atctgggctc tcggacatcg
     1681 gagaaggtga agaacaagat cttggagctc ctctacagct ggacagtggg cctgcccgag
     1741 gaggtgaaaa tcgcagaggc ctaccagatg ctaaagaagc aggggattgt aaagtccgac
     1801 cccaagcttc cagatgacac tacctttccc cttcctcctc cacggccgaa gaatgtgatc
     1861 tttgaagatg aggagaaatc caagatgctg gcccgcctgc tgaagagctc ccatcccgaa
     1921 gacctccgcg cagccaataa gctcatcaaa gagatggtgc aggaggacca gaagcggatg
     1981 gagaagatct cgaagagggt gaatgccatc gaggaggtga acaacaatgt gaaactgctc
     2041 acggagatgg tgatgagcca cagccagggc ggcgcagcag ctggcagcag cgaggacctc
     2101 atgaaggaac tgtaccagcg ctgtgagcgg atgcggccca cgctcttccg actggcgagt
     2161 gacacagagg acaatgatga ggccttagcg gagatcctgc aggccaatga caacctcacc
     2221 caggtgatca acctgtataa gcagctggtg cggggtgagg aggtcaacgg tgatgccaca
     2281 gccggctcca tccctgggag cacctcggcc ctgctggatc tctcaggcct ggatctcccg
     2341 cctgcgggca ccacctaccc agctatgccc acccgccctg gcgagcaggc cagccctgag
     2401 cagcccagtg cctcagtttc cctgcttgac gacgagctca tgtctctggg cctcagtgac
     2461 cccacacccc cttcaggccc aagcctggat ggtaccggat ggaacagctt ccagtcgtcg
     2521 gatgccactg agcccccagc ccctgctctg gcccaggccc ccagtatgga aagccgaccc
     2581 ccagcgcaga catccctgcc agcaagcagc ggtctggacg acctagacct cctggggaag
     2641 accctcctgc agcagttgct gcccccggaa tcccagcaag tgcggtggga gaagcagcag
     2701 ccaacccccc ggctcacact ccgggacctg cagaataaga gcagcagctg cagctccccc
     2761 agctccagcg ccaccagcct tctccacacc gtgtccccag agccccccag gcctccgcag
     2821 cagcccgtac caaccgagct ctcactggcc agcatcactg tgcccctgga gtccatcaaa
     2881 cccagcaaca tcctgcccgt gactgtgtat gaccagcacg gcttccgcat cctcttccat
     2941 tttgcccggg acccactgcc agggcgctcc gacgtgctgg tggtggtggt ttccatgctg
     3001 agcaccgccc cccagcccat ccgcaacatc gtgttccagt cagctgtccc caaggttatg
     3061 aaggtgaagc tgcagccacc ctcgggcacg gagctgccag cttttaaccc catcgtccac
     3121 ccctcagcaa tcacccaggt cctgctgctt gccaaccccc agaaggagaa ggttcgcctc
     3181 cgctacaagc tcaccttcac catgggtgac cagacctaca acgagatggg ggatgtggac
     3241 cagttccccc cacctgaaac ctggggtagc ctctgagtcg acggtaccgc gggcccggga
     3301 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3361 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3421 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3541 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3601 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3661 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3721 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3781 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3841 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3901 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3961 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4141 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4561 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4621 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4681 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4741 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4801 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4861 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4921 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4981 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5041 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5101 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5161 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5221 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5281 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5341 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5401 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5461 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5521 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5581 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5641 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5701 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5761 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5821 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5881 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5941 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6001 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6061 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6121 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6181 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6241 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6301 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6361 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6421 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6481 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6541 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6601 tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
