LOCUS       sequence-419581-        6595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  biosensor for cAMP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6595)
  AUTHORS   Harada K, Ito M, Wang X, Tanaka M, Wongso D, Konno A, Hirai H, 
            Hirase H, Tsuboi T, Kitaguchi T
  TITLE     Red fluorescent protein-based cAMP indicator applicable to 
            optogenetics and in vivo imaging.
  JOURNAL   Sci Rep. 2017 Aug 4;7(1):7351. doi: 10.1038/s41598-017-07820-6.
  PUBMED    28779099
REFERENCE   2  (bases 1 to 6595)
  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: "Sci Rep."; 
            date: "2017-08-4"; volume: "7(1):7351. doi"; pages: " 
            10.1038/s41598-017-07820-6"
COMMENT     Sequence Label: Pink Flamindo
FEATURES             Location/Qualifiers
     source          1..6595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(754..771)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    760..984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1030..1458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1117..1136)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1327..1348
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1467..1487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1472..1801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1652..1787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1714..1733
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1868..2662
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1922..1941)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2532..2551
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2836..2957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2873..2892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2927..2946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3006..3022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3006..3022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3019..3041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3030..3046
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3054..3060,3061..3078,3079..3084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3099..3120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3237..3254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3408..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3488..3507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4167..4958,4959..5027))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4790..4809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5028..5132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5207..5226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5398..5777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5778..5981
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5932..5952
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6026..6045
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6026..6044
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     6562..6581
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 cagtggcgcg atcctcctct cctgtcccag cgagggcctg atgccctact caccgtggcg
       61 ctccggaagc ccccaggtca gcgtacggat gaagaactgg acctgatctt tgaagagctg
      121 ctgcatatca aggcggtggc acacctttct aactcggtga agcgggaact agctgctgtt
      181 ctgctctttg aaccacacag caaggcagga actgtgttgt tcagccaggg ggacaagggt
      241 acctcgtggt acattatctg gaagggatct gtcaatgtgg tgacccatgg caaggggctg
      301 gtgaccacgt tgcacgaggg agatgacttt ggacagctgg ctctggtgaa cgacgcacct
      361 cgggcagcca ccatcatcct tcgagaaaat aactgtcact ttctgcgtgt ggacaagcag
      421 gacttcaacc gcatcatcaa ggatgtggaa tttaaagtgg agactgttga tgagcggatg
      481 taccccgagg acggcgccct gaagagcgag atcaagaagg ggctgaggct gaaggacggc
      541 ggccactacg ccgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
      601 ggcgcctaca tcgtcgacat caagttggac atcgtgtccc acaacgagga ctacaccatc
      661 gtggaacagt gcgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
      721 aagtaaaaag cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc
      781 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
      841 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
      901 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
      961 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1021 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1081 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1141 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1201 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1261 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1321 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1381 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1441 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1501 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1561 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1621 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     1681 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1741 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1801 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     1861 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     1921 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     1981 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     2041 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     2101 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     2161 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     2221 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     2281 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     2341 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     2401 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2461 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2521 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2581 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2641 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2701 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2761 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2821 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2881 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2941 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3001 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3061 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3121 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5161 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     5221 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     5281 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     5341 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
     5401 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     5461 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5521 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5581 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5641 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5701 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5761 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5821 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5881 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5941 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
     6001 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
     6061 agcgtttaaa cgggccctct agactcgagc ggccgccact gtgctggata tctgcagaat
     6121 tccaccacac tggactagtg gatccatgct ggtgagcaag ggcgaggagg ataacatggc
     6181 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     6241 gttcgagatc gagggcgagg gcgagggccg cccctacgag gcctttcaga ccgctaagct
     6301 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     6361 gtacggctcc aaggcctaca ttaagcaccc agccgacatc cccgactact tcaagctgtc
     6421 cttccccgag ggcttcaggt gggagcgcgt gatgaacttc gaggacggcg gcattattca
     6481 cgttaaccag gactcctccc tgcaggacgg cgtattcatc tacaaggtga agctgcgcgg
     6541 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggag
//
