LOCUS       sequence-487461-        9371 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  pMPS37; ubiquitously expresses cAMP sensor; zebrafish transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9371)
  AUTHORS   Sidoli M, Chen LC, Lu AJ, Wandless TJ, Talbot WS
  TITLE     A cAMP Sensor Based on Ligand-Dependent Protein Stabilization.
  JOURNAL   ACS Chem Biol. 2022 Aug 19;17(8):2024-2030. doi: 
            10.1021/acschembio.2c00333. Epub 2022 Jul 15.
  PUBMED    35839076
REFERENCE   2  (bases 1 to 9371)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acschembio.2c00333"; journalName: "ACS Chem Biol"; date: 
            "2022-08-19- 19"; volume: "17"; issue: "8"; pages: "2024-2030"
COMMENT     Sequence Label: pTol2_Ubi_N41 CNBD AcGFP
FEATURES             Location/Qualifiers
     source          1..9371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    4657..4681
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5143..5859
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     polyA_signal    5897..6031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6045..6069)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6071..6205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(6268..6288)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(6296..6312)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6328..6462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7134..7150)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7134..7150)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(7187..7207)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7187..7205)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7226..7242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7239..7261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7250..7266
                     /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(7274..7280,7281..7298,7299..7304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7319..7340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7457..7474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7628..8216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7708..7727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8387..9178,9179..9247))
                     /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     9010..9029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9248..9352)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 atgctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttgctg gatcccatcg attcgaattc
     1141 aaggcctcac cagcaaagtt ctagaatttg tcgaaacatt tatgttatat atttcctgaa
     1201 aaaaattctg agtaagttct taagtgttat tgccagcaac ataaacaaca gacggcaaaa
     1261 tgaataaatg ataacaaagc agtaggctta aataaaccta atttttatag gctgttctct
     1321 acaaccctca aacagtgatt agttttgtac ttataaactt gccctttcat tcatatttca
     1381 agaaaattgg ttcagaagat ctggatattc tagcagttgt tcaagctcat ggagggatca
     1441 gtgacctgat tccacaatga ctaggcctaa tccagaaatt agatgactgt caacataaaa
     1501 aggcacagca ctcactagct gccctatata ttttattata ttttacatat attattttat
     1561 ttatttagct ctgagtgctg tactttctgg ttaaagaaaa ctgcttacaa cagctaacct
     1621 gtactacctc aggctcaggg aatttggaac aggtttgtct ggtttgtttc tttaaccatg
     1681 catgcttgtt ttcaactatg gcaacacagt cacatgggac attacagaaa tgatttgtcg
     1741 atgacatgcg acttttcttt aataaagcgc aaagatccca aaaagcaaac ttttaacaaa
     1801 aatcatataa ttatattttc aatccagctt tgtagcaact ttgtgctgct gttcactcag
     1861 caacagatag tcagtataag gtcagtgtgt ctcaaagcag tgccatctgt ttcacacatt
     1921 gcgttctata tataagtgtg ctggttgaca cgacactgta taaggcctag gctaaaacac
     1981 aaacaatgta gaatgacact gtgttttttt tgtaaacaaa tgttgttttt ggttaaacat
     2041 ctttgtgaaa acatcctcct gtcatgtatt tgctatattc aaatgttaaa cccgtgcaga
     2101 atagaacata tacaaaaaaa aacaacacaa cacattttta aacattatta aatatcaagt
     2161 attgctggca gttctgtttc tgttttacag taccctttgc cacagttctc cgcttttcct
     2221 ggtccagatt ccacaagtct gattcaccaa tagcaaagcg aataaacaac caaagcagcc
     2281 aatcactgct tgtagactgt cctgcgagac cggcccattc cagcacattc tggaaacttc
     2341 ctttatatga taattataaa tacatttaaa ttattgatac aaaacatgta attcctagaa
     2401 cataaccata gcaatcatta gttttcaggg taattatgta tttttaggat ttgactgcgg
     2461 aaagatctgg tcatgtgacg tctcatgaac gtcacggccc tgggtttcta taaatacagt
     2521 aggactctcg accatcggca gatttttcga agaagaagat cagtttcagg agccgtactg
     2581 ttccgtttca acgcaaatat taacggtaag agcgaatttc ctagtttgtt ttcatgccat
     2641 tctttaaaac catagcgtat tactttaatt atagtaaact ttcgctttct ttattacaag
     2701 agacgttttg tgttgattct ccgcggacat tttcggtcag acaatcagaa aatgaccgcg
     2761 gaggaccagt aacttgcatt acacgtaagt taaatcttcg tgtattaaaa tggttaggtt
     2821 gttaacgtca aataggttac cgtgtttgcg tgtgatcagg ttggttttgt tagatttttg
     2881 tcagtatttt taatttattt gttttagttt atttattttt tttgctgaat catagtttgt
     2941 gaacaaagaa cccggatgtt acatacagta cagccgccat gttacagaga gttataactt
     3001 aatcatttta aaaataattt tgccttactt ttagtttgtc atgttgagaa atgaggaaat
     3061 gttaaaatga ggaaatatcc aattaattta atatatcaaa ataatccatg attacaatgc
     3121 actgaactgg agaaaattaa gatgttttct agtgtcatga aacaaatgta agagatgtac
     3181 attgtagatg ttttatgtca agaattggct agttgatgca gcatactggc gatactcagt
     3241 tgtaataaca gtaacgttac atgttaatag actactgagt atgctgttct gtctatgtat
     3301 gctctgtaag ctacgagaag gactttttta aacagtaaag ggtgcaatat ttttacaaat
     3361 tgaattaaat aaaggctgtc tattaagtaa tatgcttgat atttttctta cttgatcgga
     3421 aataagaaaa aatataaacg ttgttgctct aaaaatccta gttcagttta gccaaccaca
     3481 aatacctttt tgttcctcca acagtttttt tttcttctct ataatatttg gcagtctata
     3541 gtactccaaa tgtttcccca cagtctaact aattggtaca gccaaaatca tgacacttat
     3601 tgcaataata attttggttc attggcattg ttgatagcct gtgccactaa tatggtcgat
     3661 tgatcatgct tcaggaagaa aactatattg tttgatgtaa gattattaaa tcttcacctg
     3721 cttccattac aaactattcc catcttattg aattctggta tgtcttaaag gattagttca
     3781 cttcccaaat caaaatttac ttagcctttt ttcatccatg atcccttttt ttcatcatta
     3841 atgaagaaat tgtttttgaa aaagtttcaa gatttttttc tctatattgt ggagcttgtt
     3901 agtttaaaat tccaaaatgc aatatgtggc ttcaaatggt tctaaatgat cccagtcaag
     3961 gaataacagt cttatctaat gaaaccatta gacctttaaa aataaaaaaa ataaaagtat
     4021 ttatttttaa atgactgagt gattaagttg aatttcagcg tttccttact gtgtagaagt
     4081 ccttccttac tggccccacc ctttggttct ctgccaatct gctacctaat gtaatgttgt
     4141 ggaacattat tattctttat ttcttaattt tttatttttt attttaaaac aatgtaaact
     4201 gcacagatgt gcagtttgtt taaaatggcc aatgctttgg aaatgcatga cataattaga
     4261 tttcatgatg cacaaagcca aatctcagag cttgtgcaaa atgagctatc atttcactag
     4321 gtaagaccct aaattttcat ataggatcat ttggacaatt ttgctgcagg taaaatgcat
     4381 tctatagtcc actgtcagcc attgttttgg atagtattta tttttctcta caagtatagt
     4441 caatagtttt ctattatttt aaaggtttgt aacatttaag ggtgaccaaa tgcaaagtaa
     4501 aatttcattt tcgggtgaac tatctcgttt aacatgggag aagtgcaaaa catacattat
     4561 tggctagaac attgtagtat tttttaaatg gaaatgtgtg attgctaatc ttactttgaa
     4621 tttgtttaca gtcgagcctc tagaaccggt ggtaccgcaa gtttgtacaa aaaagcaggc
     4681 tccgaattcg cccttggatc cgccaccatg ttctaccagg aggtgaggag gggcgacttc
     4741 atgaggaact ggcagctggt ggccgccgtg cctctgttcc agaagctggg ccctgccgtg
     4801 ctggtggaga tcgtgagggc cctgagggcc aggaccgtgc ctgccggcgc cgtgatctgc
     4861 aggatcggcg agtctggcga caggatgttc ttcgtggtgg agggcagcgt gagcgtggcc
     4921 acccctaacc ctgtggagct gggccctggc gccttcttcg gcgagatggc cctgatcagc
     4981 ggcgagccta ggagcgccac cgtgagcgcc gccaccaccg tgagcctgct gagcctgcac
     5041 agcgccgact tccagatgct gtgcagcagc agccctgaga tcgccgagat catcaggaag
     5101 accgccctgg agaggagagg cgctgccgct aacgccgaat tcgtgagcaa gggcgccgag
     5161 ctgttcaccg gcatcgtgcc catcctgatc gagctgaatg gcgatgtgaa tggccacaag
     5221 ttcagcgtga gcggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     5281 atctgcacca ccggcaagct gcctgtgccc tggcccaccc tggtgaccac cctgagctac
     5341 ggcgtgcagt gcttctcacg ctaccccgat cacatgaagc agcacgactt cttcaagagc
     5401 gccatgcctg agggctacat ccaggagcgc accatcttct tcgaggatga cggcaactac
     5461 aagtcgcgcg ccgaggtgaa gttcgagggc gataccctgg tgaatcgcat cgagctgacc
     5521 ggcaccgatt tcaaggagga tggcaacatc ctgggcaata agatggagta caactacaac
     5581 gcccacaatg tgtacatcat gaccgacaag gccaagaatg gcatcaaggt gaacttcaag
     5641 atccgccaca acatcgagga tggcagcgtg cagctggccg accactacca gcagaatacc
     5701 cccatcggcg atggccctgt gctgctgccc gataaccact acctgtccac ccagagcgcc
     5761 ctgtccaagg accccaacga gaagcgcgat cacatgatct acttcggctt cgtgaccgcc
     5821 gccgccatca cccacggcat ggatgagctg tacaagtgac tcgagcctct agaactatag
     5881 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     5941 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     6001 tggaaccatt ataagctgca ataaacaagt taagggcgaa ttcgacccag ctttcttgta
     6061 caaagtgggg gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6121 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6181 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6241 tcagggggag gtgtgggagg ttttttccaa ctttattata catagttgat aattcactgg
     6301 ccgtcgtttt acggtaccat cgatgatgat ccagacatga taagatacat tgatgagttt
     6361 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     6421 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     6481 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     6541 tacaaatgtg gtatggctga ttatgatcct ctagatcaga tctgcgaaga tacggccacg
     6601 ggtgctcttg atcctgtggc tgattttgga ctgtgctgct cgcagctgct gatgaatcac
     6661 atacttcctc cattttcttc cactgattga ctgttataat ttccctaatt tccaggtcaa
     6721 ggtgctgtgc attgtggtaa tagatgtgac atgacgtcac ttccaaagga ccaatgaaca
     6781 tgtctgacca atttcatata atgtgaaaac gattttcata ggcagaataa ataacattta
     6841 aattaaactg ggcatcagcg caattcaatt ggtttggtaa tagcaaggga aaatagaatg
     6901 aagtgatctc caaaaaataa gtactttttg actgtaaata aaattgtaag gagtaaaaag
     6961 tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa agtattgatt tttaattgta
     7021 ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta cagtaatcaa gtaaaattac
     7081 tcaagtactt tacacctctg gttcttgacc ccctaccttc agcaagccca gcagatccac
     7141 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     7201 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     7801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     7861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     7921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     7981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8041 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggca
     8281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     8761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     8821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     8881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     8941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9361 cccgaaaagt g
//
