LOCUS       sequence-313705-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2P-activatable bPAC for AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Kinjo T, Watabe T, Kobachi K, Terai K, Matsuda M
  TITLE     Single-Cell Activation of the cAMP-Signaling Pathway in 3D Tissues 
            with FRET-Assisted Two-Photon Activation of bPAC.
  JOURNAL   ACS Chem Biol. 2020 Oct 19. doi: 10.1021/acschembio.0c00333.
  PUBMED    33074647
REFERENCE   2  (bases 1 to 7084)
  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: "ACS Chem 
            Biol. 2020 Oct 19. doi: 10.1021/acschembio.0c00333."
COMMENT     Sequence Label: pAAV_2pabPAC
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     705..729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          857..1235
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     CDS             3044..3100
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3101..3811
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    3826..4302
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4025..4044)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4340..4469
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4531..5119)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4611..4630)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5290..6081,6082..6150))
                     /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     5913..5932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6151..6255)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6323..6341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6379..6401)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6501..6520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6537..6992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6674..6695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6886..6905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(7067..7084,1..123)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(7078..7084,1..123)
                     /label=AAV2 ITR
ORIGIN
        1 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
       61 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      121 cctgcggcca agcgtggagc tagttattaa tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
      721 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
      781 cgatccagcc tccgcggatt cgaatcccgg ccgggaacgg tgcattggaa cgcggattcc
      841 ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccacaa aaaatgcttt
      901 cttcttttaa tatacttttt tgtttatctt atttctaata ctttccctaa tctctttctt
      961 tcagggcaat aatgatacaa tgtatcatgc ctctttgcac cattctaaag aataacagtg
     1021 ataatttctg ggttaaggca atagcaatat ttctgcatat aaatatttct gcatataaat
     1081 tgtaactgat gtaagaggtt tcatattgct aatagcagct acaatccagc taccattctg
     1141 cttttatttt atggttggga taaggctgga ttattctgag tccaagctag gcccttttgc
     1201 taatcatgtt catacctctt atcttcctcc cacagctcct gggcaacgtg ctggtctgtg
     1261 tgctggccca tcactttggc aaagaattgg gattcgaaca tcgattgaat tcaccatgat
     1321 ggtgagcaag ggcgaggaga ccacaatggg cgtaatcaag cccgacatga agatcaagct
     1381 gaagatggag ggcaacgtga atggccacgc cttcgtgatc gagggcgagg gcgagggcaa
     1441 gccctacgac ggcaccaaca ccatcaacct ggaggtgaag gagggagccc ccctgccctt
     1501 ctcctacgac attctgacca ccgcgttcgc ctacggcaac agggccttca ccaagtaccc
     1561 cgacgacatc cccaactact tcaagcagtc cttccccgag ggctactctt gggagcgcac
     1621 catgaccttc gaggacaagg gcatcgtgaa ggtgaagtcc gacatctcca tggaggagga
     1681 ctccttcatc tacgagatac acctcaaggg cgagaacttc ccccccaacg gccccgtgat
     1741 gcagaagaag accaccggct gggacgcctc caccgagagg atgtacgtgc gcgacggcgt
     1801 gctgaagggc gacgtcaagc acaagctgct gctggagggc ggcggccacc accgcgttga
     1861 cttcaagacc atctacaggg ccaagaaggc ggtgaagctg cccgactatc actttgtgga
     1921 ccaccgcatc gagatcctga accacgacaa ggactacaac aaggtgaccg tttacgagag
     1981 cgccgtggcc cgcatgatga agcggctggt gtacatcagc aagatcagcg gccacctgag
     2041 cctggaagag atccagcgga tcggcaaggt gtccatcaag aacaaccagc gggacaacat
     2101 caccggcgtg ctgctgtacc tgcagggcct gttcttccag atcctggaag gcgagaacga
     2161 gaaggtggac aagctgtaca agaaaatcct ggtggacgac cggcacacca acatcctgtg
     2221 cctgaaaacc gagtacgaca tcaccgaccg gatgttcccc aactgggcca tgaaaaccat
     2281 caacctgaac gagaacagcg agctgatgat ccagcccatc aagagcctgc tgcagaccat
     2341 cacccagagc caccgggtgc tggaaaagta catgcccgcc agagtgatct acctgatcaa
     2401 ccagggcatc aaccccctga ccgtggagcc ccagctggtc gagaagatca tcttcttcag
     2461 cgacatcctg gccttcagca ccctgaccga gaagctgccc gtgaacgagg tggtcatcct
     2521 ggtcaaccgg tacttcagca tctgcacccg gatcatcagc gcctacggcg gcgaagtgac
     2581 caagttcatc ggcgactgcg tgatggccag cttcaccaaa gagcagggcg acgccgccat
     2641 ccggaccagc ctggacatca tctccgagct gaagcagctg cggcaccacg tggaggccac
     2701 caaccccctg cacctgctgt acaccggcat cggcctgagc tacggccacg tgatcgaggg
     2761 caacatgggc agcagcctga agatggacca caccctgctg ggggacgccg tgaacgtggc
     2821 cgccaggctg gaagccctga caagacagct gccctacgcc ctggccttta cagccggcgt
     2881 gaagaagtgc tgccaggccc agtggacctt catcaacctg ggagcccacc aggtcaaggg
     2941 caagcaggaa gccatcgagg tctacaccgt gaatgaggcc cagaagtact acgacaccct
     3001 gcagatcacc cagctgatcc ggcagaccct ggaaaacgac aaggcgacga attttagtct
     3061 actgaaacaa gcgggagacg tggaggaaaa ccctggacct atggtgagca agggcgagga
     3121 gaccacaatg ggcgtaatca agcccgacat gaagatcaag ctgaagatgg agggcaacgt
     3181 gaatggccac gccttcgtga tcgagggcga gggcgagggc aagccctacg acggcaccaa
     3241 caccatcaac ctggaggtga aggagggagc ccccctgccc ttctcctacg acattctgac
     3301 caccgcgttc gcctacggca acagggcctt caccaagtac cccgacgaca tccccaacta
     3361 cttcaagcag tccttccccg agggctactc ttgggagcgc accatgacct tcgaggacaa
     3421 gggcatcgtg aaggtgaagt ccgacatctc catggaggag gactccttca tctacgagat
     3481 acacctcaag ggcgagaact tcccccccaa cggccccgtg atgcagaaga agaccaccgg
     3541 ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa
     3601 gcacaagctg ctgctggagg gcggcggcca ccaccgcgtt gacttcaaga ccatctacag
     3661 ggccaagaag gcggtgaagc tgcccgacta tcactttgtg gaccaccgca tcgagatcct
     3721 gaaccacgac aaggactaca acaaggtgac cgtttacgag agcgccgtgg cccgcaactc
     3781 caccgacggc atggacgagc tgtacaagta actcgagaga tctacgggtg gcatccctgt
     3841 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     3901 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     3961 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     4021 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     4081 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     4141 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4201 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4261 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     4321 acgtgcggac cgaggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4381 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt
     4441 gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag
     4501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6301 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6361 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6421 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6481 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     6541 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     6601 taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga
     6661 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     6721 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     6781 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     6841 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     6901 aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     6961 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt
     7021 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcccctg caggcagctg
     7081 cgcg
//
