LOCUS       sequence-473593-        5469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5469)
  AUTHORS   Kuchay S, Wang H, Marzio A, Jain K, Homer H, Fehrenbacher N, Philips
            MR, Zheng N, Pagano M
  TITLE     GGTase3 is a newly identified geranylgeranyltransferase targeting a 
            ubiquitin ligase.
  JOURNAL   Nat Struct Mol Biol. 2019 Jul;26(7):628-636. doi: 
            10.1038/s41594-019-0249-3. Epub 2019 Jun 17.
  PUBMED    31209342
REFERENCE   2  (bases 1 to 5469)
  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: 
            "10.1038/s41594-019-0249-3"; journalName: "Nat Struct Mol Biol"; 
            date: "2019-07"; volume: "26"; issue: "7"; pages: "628-636"
COMMENT     Sequence Label: pEGFPc1 Rab34
FEATURES             Location/Qualifiers
     source          1..5469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(548..1258,1259..1261,1262..1264))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(548..1258,1259..1261,1262..1264))
                     /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(590..611)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     939..958
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1198..1219
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1261..1270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1309..1512)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1338..1358)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1513..1816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1991..2579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2071..2090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2908..2955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2980..2999
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3187..3981)
                     /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             complement(3187..3981)
                     /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(3298..3317)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3908..3927
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4016..4373)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4030..4165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4182..4373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4330..4350
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4375..4479)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4506..4961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4593..4612)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4803..4824
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4968..5089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4979..4998)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5033..5052
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aatactgagc aggagtactc aggtccttct tggaacccac aaggaagagg agcacgttgg
       61 aagggtcgtt ctccttgagg gcatcagtga gccactgctt ggtgtgttcc agggatgcca
      121 cgtcattcag gttgaagaca atgatgatgg cttgagctcc acggtagtag gtggaagcaa
      181 tgcacttgaa cctttcctga ccagccgtgt cccaaagttg gagactgaag gggacaccca
      241 agacttcaaa tcgttccatc tcaaaatcca ctccgatggt agccttgtaa ttcttatcga
      301 aggtgtcttt gcagaaccta ttaatgagac aggtcttccc cacagatagg tctcccacaa
      361 cgatgacctt ggatatctta aatcccacgg tgcctgtccg gtgctcctgg caggcgcaag
      421 tgacgcgggg gtggaagtct ttgcgcacgt gcaaagcggc ctctttcttc aggcactggg
      481 gcagctccgc caggacgcgg tccctccgca cgggcgccag aatgttcact cgagatctga
      541 gtccggactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
      601 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
      661 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
      721 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
      781 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
      841 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
      901 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
      961 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     1021 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     1081 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     1141 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     1201 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     1261 ccatggtggc gaccggtagc gctagcggat ctgacggttc actaaaccag ctctgcttat
     1321 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
     1381 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
     1441 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
     1501 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     1561 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     1621 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     1681 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     1741 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     1801 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     1861 ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca ggggataacg
     1921 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1981 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2041 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2101 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2161 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2221 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2281 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2341 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2401 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2461 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2521 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2581 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2641 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2701 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     2761 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     2821 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     2881 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     2941 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     3001 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3061 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     3121 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     3181 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     3241 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     3301 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     3361 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     3421 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     3481 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     3541 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     3601 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     3661 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     3721 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     3781 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     3841 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     3901 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     3961 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc
     4021 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     4081 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     4141 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     4201 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     4261 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4321 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt
     4381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4501 acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4561 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4621 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4681 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4741 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4801 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4861 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4921 atttaacgcg aattttaaca aaatattaac gcttacaatt tacgcgttaa gatacattga
     4981 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5041 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5101 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     5161 aaacctctac aaatgtggta tggctgatta tgatcagtta tctagatccg gtggatcctc
     5221 agggacaaca tgtggccttc ttcttgctgg cagttaggta caggtttttg tcatcgctgt
     5281 tgatgcgaac aacatctgca atgtgccgag ccccggattt ctccagctcc gccaggacat
     5341 tggcctcaaa ggttagtgct gccacacgga agaagaattc ccggacattc tcaccagtga
     5401 gggaggacac cgcccagtac tcggccttaa tctcttgggc caccttgagg gcatccttct
     5461 ccattaggg
//
