LOCUS       sequence-485017-        4153 bp    DNA     circular SYN 29-AUG-2025
DEFINITION  pMPS53; middle entry clone holding N41_R307Q_CNBD_acgfp 
            (cAMP-insensitive mutant control sensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4153)
  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 4153)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 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: pME_N41_R307Q_CNBD_AcGFP
FEATURES             Location/Qualifiers
     source          1..4153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(8..107)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(125..143)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(148..164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(148..164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             595..1386
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1479..2067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1968..1987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2221..2238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2397..2424)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2516..2602)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2651..2673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2665..2682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2666..2682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2698..2797
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3259..3975
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     polyA_signal    4013..4147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4037..4056)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4091..4110
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gaattcgacc cagctttctt gtacaaagtt ggcattataa aaaataattg ctcatcaatt
       61 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
      121 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      181 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag
      241 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
      301 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
      361 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
      421 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
      481 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
      541 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
      601 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
      661 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
      721 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
      781 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
      841 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
      901 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
      961 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1021 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1081 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1141 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1201 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1261 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     1321 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     1381 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     1441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1621 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2101 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2161 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2221 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2281 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2341 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     2401 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     2461 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     2521 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     2581 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     2641 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa
     2701 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc
     2761 ttttttataa tgccaacttt gtacaaaaaa gcaggctccg aattcgccct tggatccgcc
     2821 accatgttct accaggaggt gaggaggggc gacttcatga ggaactggca gctggtggcc
     2881 gccgtgcctc tgttccagaa gctgggccct gccgtgctgg tggagatcgt gagggccctg
     2941 agggccagga ccgtgcctgc cggcgccgtg atctgcagga tcggcgagtc tggcgacagg
     3001 atgttcttcg tggtggaggg cagcgtgagc gtggccaccc ctaaccctgt ggagctgggc
     3061 cctggcgcct tcttcggcga gatggccctg atcagcggcg agcctcagag cgccaccgtg
     3121 agcgccgcca ccaccgtgag cctgctgagc ctgcacagcg ccgacttcca gatgctgtgc
     3181 agcagcagcc ctgagatcgc cgagatcatc aggaagaccg ccctggagag gagaggcgct
     3241 gccgctaacg ccgaattcgt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc
     3301 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag
     3361 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct
     3421 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac
     3481 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag
     3541 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc
     3601 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc
     3661 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc
     3721 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc
     3781 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg
     3841 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag
     3901 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat
     3961 gagctgtaca agtgactcga gcctctagaa ctatagtgag tcgtattacg tagatccaga
     4021 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4081 ctttatttgt gaaatttgtg atgctattgc tttatttgga accattataa gctgcaataa
     4141 acaagttaag ggc
//
