LOCUS       sequence-232214-        5253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Rap2b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5253)
  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 5253)
  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-Rap2b
FEATURES             Location/Qualifiers
     source          1..5253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        168..471
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        472..675
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     626..646
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      714..723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(720..722,723..725,726..1436)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(720..722,723..725,726..1436)
                     /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(765..786)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1026..1045)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1373..1394
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2148..2269
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2185..2204)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2239..2258
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2276..2731)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2413..2434)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2625..2644
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2758..2862
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2864..3221
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2864..3055
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2887..2907)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3072..3207
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3256..4050
                     /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             3256..4050
                     /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(3310..3329)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3920..3939
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4238..4257)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4282..4329
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4658..5246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5147..5166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
       61 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
      121 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      181 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      241 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      301 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      361 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      421 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      481 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      541 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      601 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      661 tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg gtcgccacca
      721 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
      781 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
      841 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
      901 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
      961 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1021 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1081 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1141 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1201 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1261 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1321 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1381 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     1441 gactcagatc tcgagctcaa gcttcgaatt ctatgagaga gtacaaagtg gtggtgctgg
     1501 gctcgggcgg cgtgggcaag tccgcgctca ccgtgcagtt cgtgacgggc tccttcatcg
     1561 agaagtacga cccgaccatc gaagactttt accgcaagga gattgaggtg gactcgtcgc
     1621 cgtcggtgct ggagatcctg gatacggcgg gcaccgagca gttcgcgtcc atgcgggacc
     1681 tgtacatcaa gaacggccag ggcttcatcc tggtctacag cctcgtcaac cagcagagct
     1741 tccaggacat caagcccatg cgggaccaga tcatccgcgt gaagcggtac gagcgcgtgc
     1801 ccatgatcct ggtgggcaac aaggtggacc tggagggtga gcgcgaggtc tcgtacgggg
     1861 agggcaaggc cctggctgag gagtggagct gccccttcat ggagacgtcg gccaaaaaca
     1921 aagcctcggt agacgagcta tttgccgaga tcgtgcggca gatgaactac gcggcgcagc
     1981 ccaacggcga tgagggctgc tgctcggcct gcgtgatcct ctgactcgag tctagataac
     2041 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2101 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2281 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2701 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2881 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2941 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3061 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3121 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3181 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3241 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3301 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3361 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3421 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3481 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3541 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3601 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3661 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3721 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3781 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3841 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3901 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3961 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4021 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4081 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4141 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4201 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4261 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4321 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4381 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4441 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4501 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4561 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4621 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4681 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4741 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4801 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4861 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4921 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4981 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5041 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5101 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5161 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5221 cgtcaggggg gcggagccta tggaaaaacg cca
//
