LOCUS       sequence-382898-        8269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of cAMP sensor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8269)
  TITLE     Expression of cAMP sensor
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8269)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: Epac-SH226
FEATURES             Location/Qualifiers
     source          1..8269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        137..440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        441..644
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     595..615
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(677..679,680..682,683..1195)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(722..743)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(983..1002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1330..1351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3581..5011
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     polyA_signal    5133..5254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5170..5189)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5224..5243
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5261..5716)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5398..5419)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5610..5629
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5743..5847
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5849..6206
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5849..6040
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5872..5892)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6057..6192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6241..7035
                     /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             6241..7035
                     /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(6295..6314)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6905..6924
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7223..7242)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7267..7314
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7643..8231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8132..8151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
       61 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      121 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      181 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      481 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      541 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      601 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      661 agatccgcta gccgcaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      721 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      781 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      841 cgtgccctgg cccaccctcg tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta
      901 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      961 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1021 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1081 caacatcctg gggcacaagc tggagtacaa ctactttagc gacaacgtct atatcaccgc
     1141 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
     1201 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1261 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     1321 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc cgatatcagc ccgtgggaac
     1381 tcatgagatg gaggaggagt tggccgaagc tgtggccctg ctctcccagc gggggcctga
     1441 cgccctgctc actgtggcac ttcgaaagcc cccaggtcag cgcacggatg aagagctgga
     1501 cctcatcttt gaggagctgc tgcacatcaa ggctgtggcc cacctctcca actcggtgaa
     1561 gcgagaatta gcggctgttc tgctctttga accacacagc aaggcaggga ccgtgttgtt
     1621 cagccagggg gacaagggca cttcgtggta cattatctgg aagggatctg tcaacgtggt
     1681 gacccatggc aaggggctgg tgaccaccct gcatgaggga gatgattttg gacagctggc
     1741 tctggtgaat gatgcacccc gggcagccac catcatcctg cgagaagaca actgtcattt
     1801 cctgcgtgtg gacaagcagg acttcaaccg tatcatcaag gatgtggagg caaagaccat
     1861 gcggctggaa gaacatggca aagtggtgct ggtgctggag agagcctctc agggcgccgg
     1921 cccttcccga cccccaaccc caggcaggaa ccggtataca gtgatgtctg gcaccccaga
     1981 gaagatccta gagcttctgt tggaggccat gggaccagat tccagtgctc atgacccaac
     2041 agagacattc ctcagcgact tcctcctgac ccacagggtc ttcatgccca gcgcccaact
     2101 ctgcgctgcc cttctgcacc acttccatgt ggagcctgcg ggtggcagcg agcaggagcg
     2161 cagcacctac gtctgcaaca agaggcagca gatcttgcgg ctggtcagcc agtgggtggc
     2221 cctgtatggc tccatgctcc acactgaccc tgtggccacc agcttcctcc agaaactctc
     2281 agacctggtg ggcagggaca cccgactcag caacctgctg agggagcagt ggccagagag
     2341 gcggcgatgc cacaggttgg agaatggctg tgggaatgca tctcctcaga tgaaggcccg
     2401 gaacttgcct gtttggctcc ccaaccagga cgagcccctt cctggcagca gctgtgccat
     2461 ccaagttggg gataaagtcc cctatgacat ctgccggcca gaccactcag tgttgaccct
     2521 gcagctgcct gtgacagcct ccgtgagaga ggtgatggca gcgttggccc aggaggatgg
     2581 ctggaccaag gggcaggtgc tggtgaaggt caattctgca ggtgatgcca ttggcctgca
     2641 gccagatgcc cgtggtgtgg ccacatctct ggggctcaat gagcgtctct ttgttgtcaa
     2701 cccacaggaa gcgcatgagc tgatcccaca ccctgaccag ctggggccca ctgtgggctc
     2761 tgctgagggg ctggacctgg tgagtgccaa ggacctggca ggccagctga cggaccacga
     2821 ctggagcctc ttcaacagta tccaccaggt ggagctgatc cactatgtgc tgggccccca
     2881 gcatctgcgg gatgtcacca ccgccaacct ggagcgcttc atgcgccgct tcaatgagct
     2941 gcagtactgg gtggccaccg agctgtgtct ctgccccgtg cccggccccc gggcccagct
     3001 gctcaggaag ttcattaagc tggcggccca cctcaaggag cagaagaatc tcaattcctt
     3061 ctttgccgtc atgtttggcc tcagcaactc ggccatcagc cgcctagccc acacctggga
     3121 gcggctgccc cacaaagtcc ggaagctgta ctccgccctc gagaggctgc tggatccctc
     3181 atggaaccac cgggtatacc gactggccct cgccaagctc tcccctcctg tcatcccctt
     3241 catgcccctt cttctcaaag acatggccgc cattcatgag ggaaaccaca cactagtgga
     3301 gaatctcatc aactttgaga agatgagaat gatggccaga gccgcgcgga tgctgcacca
     3361 ctgccgaagc cacaaccctg tgcctctctc accactcaga agccgagttt cccacctcca
     3421 cgaggacagc caggtggcga ggatttccac atgctcggag cagtccctga gcacccggag
     3481 tccagccagc acctgggctt atgtccagca gctgaaggtc attgacaacc agcgggaact
     3541 ctcccgcctc tcccgagagc tggagccagc aagcgagctc atggtgagca agggcgagga
     3601 ggtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccatga acggccacga
     3661 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3721 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcat
     3781 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgattaca agaagctgtc
     3841 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gtctggtgac
     3901 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agatgcgcgg
     3961 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     4021 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     4081 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     4141 cgtgcaactg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     4201 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctggg
     4261 gcatggcacc ggcagcaccg gcagcggcag ctccggcacc gcctcctccg aggacaacaa
     4321 catggccgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccatgaacgg
     4381 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     4441 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     4501 gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg attacaagaa
     4561 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggtct
     4621 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagat
     4681 gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4741 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     4801 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     4861 gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     4921 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     4981 cctgtacggc atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc
     5041 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5101 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5161 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5221 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     5281 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5341 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     5401 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     5461 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     5521 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     5581 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     5641 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     5701 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5761 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5821 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     5881 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5941 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     6001 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6061 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6121 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6181 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     6241 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     6301 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     6361 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     6421 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     6481 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     6541 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     6601 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     6661 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     6721 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     6781 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     6841 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     6901 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     6961 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7021 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     7081 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     7141 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     7201 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     7261 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     7321 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     7381 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     7441 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     7501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggcctt
//
