LOCUS       sequence-227882-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Ran.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  AUTHORS   Liu C, Takahashi M, Li Y, Dillon TJ, Kaech S, Stork PJ
  TITLE     The interaction of Epac1 and Ran promotes Rap1 activation at the 
            nuclear envelope.
  JOURNAL   Mol Cell Biol. 2010 Aug;30(16):3956-69. doi: 10.1128/MCB.00242-10. 
            Epub 2010 Jun 14.
  PUBMED    20547757
REFERENCE   2  (bases 1 to 7986)
  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: 
            "10.1128/MCB.00242-10"; journalName: "Mol Cell Biol"; date: 
            "2010-08"; volume: "30"; issue: "16"; pages: "3956-69"
COMMENT     Sequence Label: GFP-RanV19 E1RA2
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        131..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      677..686
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(683..685,686..688,689..1399)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(683..685,686..688,689..1399)
                     /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(728..749)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(989..1008)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1336..1357
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4844..4965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4881..4900)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4935..4954
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4972..5427)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5109..5130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5321..5340
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5454..5558
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5560..5917
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5560..5751
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5583..5603)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5768..5903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5952..6746
                     /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             5952..6746
                     /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(6006..6025)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6616..6635
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6934..6953)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6978..7025
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7354..7942
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7843..7862
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
       61 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      661 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      721 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      781 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      841 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      901 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      961 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1021 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1081 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     1141 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     1201 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1261 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1321 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1381 catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga tggctgcgca
     1441 gggagagccc caggtccagt tcaaacttgt attggttggt gatgttggta ctggaaaaac
     1501 gaccttcgtg aaacgtcatt tgactggtga atttgagaag aagtatgtag ccaccttggg
     1561 tgttgaggtt catcccctag tgttccacac caacagagga cctattaagt tcaatgtatg
     1621 ggacacagcc ggccaggaga aattcggtgg actgagagat ggctattata tccaagccca
     1681 gtgtgccatc ataatgtttg atgtaacatc gagagttact tacaagaatg tgcctaactg
     1741 gcatagagat ctggtacgag tgtgtgaaaa catccccatt gtgttgtgtg gcaacaaagt
     1801 ggatattaag gacaggaaag tgaaggcgaa atccattgtc ttccaccgaa agaagaatct
     1861 tcagtactac gacatttctg ccaaaagtaa ctacaacttt gaaaagccct tcctctggct
     1921 tgctaggaag ctcattggag accctaactt ggaatttgtt gccatgcctg ctctcgcccc
     1981 accagaagtt gtcatggacc cagctttggc agcacagtat gagcacgact tagaggttgc
     2041 tcagacaact gctctcccgg atgaggatga tgacctggtg aattcaatgg tgttgagaag
     2101 gatgcaccgg ccccgaagct gctcctacca gctgctgctg gagcaccagc gtccgagctg
     2161 catccagggg ctgcgctgga caccactcac caacagcgag gagtccctgg atttcagcga
     2221 gagcctggag caggcctcca cagagcgggt gctcagggct gggaggcagc tgcatcggca
     2281 tctgctggcc acctgcccaa acctcacccg agaccggaag taccacctta ggctctatcg
     2341 gcagtgctgc tctggccggg agctggtgga tgggatcttg gccctgggac ttggggtcca
     2401 ttcccggagc caagttgtgg gaatctgcca ggtgctgctg gatgaaggtg ccctctgcca
     2461 tgtgaaacac gactgggcct tccaggaccg agatgcccaa ttctaccggt tccccgggcc
     2521 cgagcccgag cccgtgggaa ctcatgagat ggaggaggag ttggccgaag ctgtggccct
     2581 gctctcccag cgggggcctg acgccctgct cactgtggca cttcgaaagc ccccaggtca
     2641 gcgcacggat gaagagctgg acctcatctt tgaggagctg ctgcacatca aggctgtggc
     2701 ccacctctcc aactcggtga agcgagaatt agcggctgtt ctgctctttg aaccacacag
     2761 caaggcaggg accgtgttgt tcagccaggg ggacaagggc acttcgtggt acattatctg
     2821 gaagggatct gtcaacgtgg tgacccatgg caaggggctg gtgaccaccc tgcatgaggg
     2881 agatgatttt ggacagctgg ctctggtgaa tgatgcaccc cgggcagcca ccatcatcct
     2941 gcgagaagac aactgtcatt tcctgcgtgt ggacaagcag gacttcaacc gtatcatcaa
     3001 ggatgtggag gcaaagacca tgcggctgga agaacatggc aaagtggtgc tggtgctgga
     3061 gagagcctct cagggcgccg gcccttcccg acccccaacc ccaggcagga accggtatac
     3121 agtgatgtct ggcaccccag agaagatcct agagcttctg ttggaggcca tgggaccaga
     3181 ttccagtgct catgacccaa cagagacatt cctcagcgac ttcctcctga cccacagggt
     3241 cttcatgccc agcgcccaac tctgcgctgc ccttctgcac cacttccatg tggagcctgc
     3301 gggtggcagc gagcaggagc gcagcaccta cgtctgcaac aagaggcagc agatcttccg
     3361 cttggtcctg cagtgggcgg ccatgtatgg cgatctcctc caagaagatg atgtggccat
     3421 ggccttcctg gaggagttct atgtgtctgt atcagatgac gcacggatga tggctgcctt
     3481 caaggagcag ctgccagagc tggagaagat tgtcaagcaa atctcagaag acgcaaaagc
     3541 tccacagaag aagcacaagg tgcttttgca acagttcaac acaggtgacg agagggccca
     3601 gaagcgtcag cctattcgtg gctctgatga ggttttgttc aaggtctact gcatcgacca
     3661 cacctatact accattcgtg tgccggtagc tgcctcggtg aaggaagtca tcagtgcagt
     3721 agctgacaaa ctgggctcag gggaaggcct gatcatcgtc aagatgaact ctggaggaga
     3781 aaaggtggtg ctgaaatcta atgatgtttc agtatttacg acgctcacca ttaataatcg
     3841 attgtttgtt gtcaacccac aggaagcgca tgagctgatc ccacaccctg accagctggg
     3901 gcccactgtg ggctctgctg aggggctgga cctggtgagt gccaaggacc tggcaggcca
     3961 gctgacggac cacgactgga gcctcttcaa cagtatccac caggtggagc tgatccacta
     4021 tgtgctgggc ccccagcatc tgcgggatgt caccaccgcc aacctggagc gcttcatgcg
     4081 ccgcttcaat gagctgcagt actgggtggc caccgagctg tgtctctgcc ccgtgcccgg
     4141 cccccgggcc cagctgctca ggaagttcat taagctggcg gcccacctca aggagcagaa
     4201 gaatctcaat tccttctttg ccgtcatgtt tggcctcagc aactcggcca tcagccgcct
     4261 agcccacacc tgggagcggc tgccccacaa agtccggaag ctgtactccg ccctcgagag
     4321 gctgctggat ccctcatgga accaccgggt ataccgactg gccctcgcca agctctcccc
     4381 tcctgtcatc cccttcatgc cccttcttct caaagacatg accttcattc atgagggaaa
     4441 ccacacacta gtggagaatc tcatcaactt tgagaagatg agaatgatgg ccagagccgc
     4501 gcggatgctg caccactgcc gaagccacaa ccctgtgcct ctctcaccac tcagaagccg
     4561 agtttcccac ctccacgagg acagccaggt ggcgaggatt tccacatgct cggagcagtc
     4621 cctgagcacc cggagtccag ccagcacctg ggcttatgtc cagcagctga aggtcattga
     4681 caaccagcgg gaactctccc gcctctcccg agagctggag ccatagtcta gataactgat
     4741 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4801 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     4981 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5041 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5101 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5161 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5221 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5281 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5341 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5401 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5461 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5521 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     5581 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5641 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     5701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     5761 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5821 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5881 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     5941 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6001 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6061 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6121 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6181 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6241 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6301 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6361 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6421 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     6481 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6541 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6601 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6661 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6721 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     6781 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6841 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6901 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6961 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     7021 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     7081 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     7141 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     7201 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7261 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7321 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7381 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7441 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7501 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7561 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     7621 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     7681 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     7741 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     7801 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     7861 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7921 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7981 ttgctg
//
