LOCUS       sequence-357440-        6330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GCP60.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6330)
  AUTHORS   Sbodio JI, Hicks SW, Simon D, Machamer CE
  TITLE     GCP60 preferentially interacts with a caspase-generated golgin-160 
            fragment.
  JOURNAL   J Biol Chem. 2006 Sep 22;281(38):27924-31. doi: 
            10.1074/jbc.M603276200. Epub 2006 Jul 26.
  PUBMED    16870622
REFERENCE   2  (bases 1 to 6330)
  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: 
            "10.1074/jbc.M603276200"; journalName: "J Biol Chem"; date: 
            "2006-09-22- 22"; volume: "281"; issue: "38"; pages: "27924-31"
COMMENT     Sequence Label: pEGFP/GCP60
FEATURES             Location/Qualifiers
     source          1..6330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        72..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     608..627
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        608..626
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(647..649,650..652,653..1363)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(647..649,650..652,653..1363)
                     /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(692..713)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(953..972)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1300..1321
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1380..1389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3129..3250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3166..3185)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3220..3239
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3257..3712)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3394..3415)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3606..3625
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3739..3843
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3845..4202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3845..4036
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3868..3888)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4053..4188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4237..5031
                     /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             4237..5031
                     /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(4291..4310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4901..4920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5219..5238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5263..5310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5639..6227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6128..6147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagctaa tacgactcac tatagggaga cccaccggtc gccaccatgg tgagcaaggg
      661 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      721 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      781 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      841 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      901 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      961 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1021 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1081 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1141 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1201 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1261 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1321 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctac
     1381 catggcggcg gtgctgaacg cagagcgact cgaggtgtcc gtcgacggcc tcacgctcag
     1441 cccggacccg gaggagcggc ctggggcgga gggcgccccg ctgctgccgc caccgctgcc
     1501 accgccctcg ccacctggat ccggtcgcgg cccgggcgcc tcaggggagc agcccgagcc
     1561 cggggaggcg gcggctgggg gcgcggcgga ggaggcgcgg cggctggagc agcgctgggg
     1621 tttcggcctg gaggagttgt acggcctggc actgcgcttc ttcaaagaaa aagatggcaa
     1681 agcatttcat ccaacttatg aagaaaaatt gaagcttgtg gcactgcata agcaagttct
     1741 tatgggccca tataatccag acacttgtcc tgaggttgga ttctttgatg tgttggggaa
     1801 tgacaggagg agagaatggg cagccctggg aaacatgtct aaagaggatg ccatggtgga
     1861 gtttgtcaag ctcttaaata ggtgttgcca tctcttttca acatatgttg cgtcccacaa
     1921 aatagagaag gaagagcaag aaaaaaaaag gaaggaggaa gaggagcgaa ggcggcgtga
     1981 agaggaagaa agagaacgtc tgcaaaagga ggaagagaaa cgtaggagag aagaagagga
     2041 aaggcttcga cgggaggaag aggaaaggag acggatagaa gaagaaaggc ttcggttgga
     2101 gcagcaaaag cagcagataa tggcagcttt aaactcccag actgccgtgc agttccagca
     2161 gtatgcagcc caacagtatc cagggaacta cgaacagcag caaattctca tccgccagtt
     2221 gcaggagcaa cactatcagc agtacatgca gcagttgtat caagtccagc ttgcacagca
     2281 acaggcagca ttacagaaac aacaggaagt agtagtggct gggtcttcct tgcctacatc
     2341 atcaaaagtg aatgcaactg taccaagtaa tatgatgtca gttaatggac aggccaaaac
     2401 acacactgac agctccgaaa aagaactgga accagaagct gcagaagaag ccctggagaa
     2461 tggaccaaaa gaatctcttc cagtaatagc agctccatcc atgtggacac gacctcagat
     2521 caaagacttc aaagagaaga ttcagcagga tgcagattcc gtgattacag tgggccgagg
     2581 agaagtggtc actgttcgag tacccaccca tgaagaagga tcatatctct tttgggaatt
     2641 tgccacagac aattatgaca ttgggtttgg ggtgtatttt gaatggacag actctccaaa
     2701 cactgctgtc agcgtgcatg tcagtgagtc cagcgatgac gacgaggagg aagaagaaaa
     2761 catcggttgt gaagagaaag ccaaaaagaa tgccaacaag cctttgctgg atgagattgt
     2821 gcctgtgtac cgacgggact gtcatgagga ggtgtatgct ggcagccatc aatatccagg
     2881 gagaggagtc tatctcctca agtttgacaa ctcctactct ttgtggcggt caaaatcagt
     2941 ctactacaga gtctattata ctagatgaga attctgcagt cgacggtacc gcgggcccgg
     3001 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     3061 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3121 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3241 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3301 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3361 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3421 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3481 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3541 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3601 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3661 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3721 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3781 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3841 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3901 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3961 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4021 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4081 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4141 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4201 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4381 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4681 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4741 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4921 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5041 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5101 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5161 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5221 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5281 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5341 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5401 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5461 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5521 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5581 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5641 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5701 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5761 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5821 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5881 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5941 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6001 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6061 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6301 tatcccctga ttctgtggat aaccgtatta
//
