LOCUS       sequence-229774-        5362 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of human Ran.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5362)
  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 5362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: mCherry-RanV19
FEATURES             Location/Qualifiers
     source          1..5362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             617..1324
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(762..780)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1030..1049
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2154..2275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2191..2210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2245..2264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2282..2737)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2631..2650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2764..2868
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2870..3227
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2870..3061
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2893..2913)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3078..3213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3262..4056
                     /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             3262..4056
                     /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(3316..3335)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3926..3945
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4244..4263)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4288..4335
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4664..5252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5153..5172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      661 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      721 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      781 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      841 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
      901 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
      961 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1021 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     1081 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1141 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1201 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     1261 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     1321 caagtccgga ctcagatctc gagctcaagc ttcgaattcc atggctgcgc agggagagcc
     1381 ccaggtccag ttcaaacttg tattggttgg tgatgttggt actggaaaaa cgaccttcgt
     1441 gaaacgtcat ttgactggtg aatttgagaa gaagtatgta gccaccttgg gtgttgaggt
     1501 tcatccccta gtgttccaca ccaacagagg acctattaag ttcaatgtat gggacacagc
     1561 cggccaggag aaattcggtg gactgagaga tggctattat atccaagccc agtgtgccat
     1621 cataatgttt gatgtaacat cgagagttac ttacaagaat gtgcctaact ggcatagaga
     1681 tctggtacga gtgtgtgaaa acatccccat tgtgttgtgt ggcaacaaag tggatattaa
     1741 ggacaggaaa gtgaaggcga aatccattgt cttccaccga aagaagaatc ttcagtacta
     1801 cgacatttct gccaaaagta actacaactt tgaaaagccc ttcctctggc ttgctaggaa
     1861 gctcattgga gaccctaact tggaatttgt tgccatgcct gctctcgccc caccagaagt
     1921 tgtcatggac ccagctttgg cagcacagta tgagcacgac ttagaggttg ctcagacaac
     1981 tgctctcccg gatgaggatg atgacctgta atctagaggg cccgggatcc accggatcta
     2041 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2101 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2161 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2221 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
     2281 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2341 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2401 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2461 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2521 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2581 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2641 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2701 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2761 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2821 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     2881 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     2941 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3001 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3061 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3121 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3181 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3241 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3301 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3361 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3421 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3481 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3541 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3601 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3661 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3721 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3781 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3841 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3901 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3961 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4021 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4081 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4141 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4201 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4261 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4321 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4381 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4441 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4501 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4561 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4621 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4681 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4741 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4801 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4861 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4921 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4981 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5041 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5101 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5161 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5221 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5281 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5341 tggataaccg tattaccgcc at
//
