LOCUS       sequence-233713-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human EPAC1 gene was fused in-frame and upstream from the enhanced 
            YFP gene in pEYFP-N3 vector (Clonetech)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Mei FC, Qiao J, Tsygankova OM, Meinkoth JL, Quilliam LA, Cheng X
  TITLE     Differential signaling of cyclic AMP: opposing effects of exchange 
            protein directly activated by cyclic AMP and cAMP-dependent protein 
            kinase on protein kinase B activation.
  JOURNAL   J Biol Chem. 2002 Mar 29;277(13):11497-504. doi: 
            10.1074/jbc.M110856200. Epub 2002 Jan 18.
  PUBMED    11801596
REFERENCE   2  (bases 1 to 7354)
  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.M110856200"; journalName: "J Biol Chem"; date: 
            "2002-03-29- 29"; volume: "277"; issue: "13"; pages: "11497-504"
COMMENT     Sequence Label: pEYFP-N3-EPAC1
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3294..3303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3300..3302,3303..3305,3306..4019)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3345..3366)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3606..3625)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3953..3974
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4142..4263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4179..4198)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4233..4252
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4270..4725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4619..4638
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4752..4856
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4858..5215
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4858..5049
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4881..4901)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5066..5201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5250..6044
                     /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             5250..6044
                     /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(5304..5323)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5914..5933
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6232..6251)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6276..6323
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6652..7240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7141..7160
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 tgttgagaag gatgcaccgg ccccgaagct gctcctacca gctgctgctg gagcaccagc
      661 gtccgagctg catccagggg ctgcgctgga caccactcac caacagcgag gagtccctgg
      721 atttcagcga gagcctggag caggcctcca cagagcgggt gctcagggct gggaggcagc
      781 tgcatcggca tctgctggcc acctgcccaa acctcacccg agaccggaag taccacctta
      841 ggctctatcg gcagtgctgc tctggccggg agctggtgga tgggatcttg gccctgggac
      901 ttggggtcca ttcccggagc caagttgtgg gaatctgcca ggtgctgctg gatgaaggtg
      961 ccctctgcca tgtgaaacac gactgggcct tccaggaccg agatgcccaa ttctaccggt
     1021 tccccgggcc cgagcccgag cccgtgggaa ctcatgagat ggaggaggag ttggccgaag
     1081 ctgtggccct gctctcccag cgggggcctg acgccctgct cactgtggca cttcgaaagc
     1141 ccccaggtca gcgcacggat gaagagctgg acctcatctt tgaggagctg ctgcacatca
     1201 aggctgtggc ccacctctcc aactcggtga agcgagaatt agcggctgtt ctgctctttg
     1261 aaccacacag caaggcaggg accgtgttgt tcagccaggg ggacaagggc acttcgtggt
     1321 acattatctg gaagggatct gtcaacgtgg tgacccatgg caaggggctg gtgaccaccc
     1381 tgcatgaggg agatgatttt ggacagctgg ctctggtgaa tgatgcaccc cgggcagcca
     1441 ccatcatcct gcgagaagac aactgtcatt tcctgcgtgt ggacaagcag gacttcaacc
     1501 gtatcatcaa ggatgtggag gcaaagacca tgcggctgga agaacatggc aaagtggtgc
     1561 tggtgctgga gagagcctct cagggcgccg gcccttcccg acccccaacc ccaggcagga
     1621 accggtatac agtgatgtct ggcaccccag agaagatcct agagcttctg ttggaggcca
     1681 tgggaccaga ttccagtgct catgacccaa cagagacatt cctcagcgac ttcctcctga
     1741 cccacagggt cttcatgccc agcgcccaac tctgcgctgc ccttctgcac cacttccatg
     1801 tggagcctgc gggtggcagc gagcaggagc gcagcaccta cgtctgcaac aagaggcagc
     1861 agatcttgcg gctggtcagc cagtgggtgg ccctgtatgg ctccatgctc cacactgacc
     1921 ctgtggccac cagcttcctc cagaaactct cagacctggt gggcagggac acccgactca
     1981 gcaacctgct gagggagcag tggccagaga ggcggcgatg ccacaggttg gagaatggct
     2041 gtgggaatgc atctcctcag atgaaggccc ggaacttgcc tgtttggctc cccaaccagg
     2101 acgagcccct tcctggcagc agctgtgcca tccaagttgg ggataaagtc ccctatgaca
     2161 tctgccggcc agaccactca gtgttgaccc tgcagctgcc tgtgacagcc tccgtgagag
     2221 aggtgatggc agcgttggcc caggaggatg gctggaccaa ggggcaggtg ctggtgaagg
     2281 tcaattctgc aggtgatgcc attggcctgc agccagatgc ccgtggtgtg gccacatctc
     2341 tggggctcaa tgagcgtctc tttgttgtca acccacagga agcgcatgag ctgatcccac
     2401 accctgacca gctggggccc actgtgggct ctgctgaggg gctggacctg gtgagtgcca
     2461 aggacctggc aggccagttg acggaccacg actggagcct cttcaacagt atccaccagg
     2521 tggagctgat ccactatgtg ctgggccccc agcatctgcg ggatgtcacc accgccaacc
     2581 tggagcgctt catgcgccgc ttcaatgagc tgcagtactg ggtggccacc gagctgtgtc
     2641 tctgccccgt gcccggcccc cgggcccagc tgctcaggaa gttcattaag ctggcggccc
     2701 acctcaagga gcagaagaat ctcaattcct tctttgccgt catgtttggc ctcagcaact
     2761 cggccatcag ccgcctagcc cacacctggg agcgactgcc ccacaaagtc cggaagctgt
     2821 actccgccct cgagaggctg ctggatccct catggaacca ccgggtatac cgactggccc
     2881 tcgccaagct ctcccctcct gtcatcccct tcatgcccct tcttctcaaa gacatgacct
     2941 tcattcatga gggaaaccac acactagtgg agaatctcat caactttgag aagatgagaa
     3001 tgatggccag agccgcgcgg atgctgcacc actgccgaag ccacaaccct gtgcctctct
     3061 caccactcag aagccgagtt tcccacctcc acgaggacag ccaggtggcg aggatttcca
     3121 catgctcgga gcagtccctg agcacccgga gtccagccag cacctgggct tatgtccagc
     3181 agctgaaggt cattgacaac cagcgggaac tctcccgcct ctcccgagag ctggagccag
     3241 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc atcgccacca
     3301 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3361 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3421 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3481 tcgtgaccac cttcggctac ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     3541 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3601 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3661 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3721 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3781 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3841 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3901 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3961 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     4021 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     4081 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     4141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4261 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4381 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4441 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4501 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4561 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4621 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4681 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     4741 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4801 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     4861 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4981 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5041 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5101 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5161 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     5221 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5281 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5341 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5401 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5461 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5521 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5581 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5641 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5701 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5761 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5821 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5881 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5941 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6001 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6061 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     6121 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6181 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     6241 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     6301 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     6361 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     6421 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     6481 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     6541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6781 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7261 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7321 tgattctgtg gataaccgta ttaccgccat gcat
//
