LOCUS       Exported                7084 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  A negative control for AAV-mediated 2P-activatable bPAC.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV_2pabPAC(Y72L)
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7084
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        23..326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        327..530
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     481..501
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     527..551
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2866..2922
                     /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             2923..3633
                     /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    3648..4124
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3847..3866)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4162..4291
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4353..4941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4433..4452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5112..5972)
                     /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     5735..5754
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5973..6077)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6145..6163
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6201..6223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6323..6342
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6359..6814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6496..6517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6708..6727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6889..7029
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6900..7029
                     /label=AAV2 ITR
ORIGIN
        1 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
       61 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      121 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      181 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      241 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      301 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      361 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      421 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      481 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      541 accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg
      601 accgatccag cctccgcgga ttcgaatccc ggccgggaac ggtgcattgg aacgcggatt
      661 ccccgtgcca agagtgacgt aagtaccgcc tatagagtct ataggcccac aaaaaatgct
      721 ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc
      781 tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag
      841 tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa
      901 attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc
      961 tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt
     1021 gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg
     1081 tgtgctggcc catcactttg gcaaagaatt gggattcgaa catcgattga attcaccatg
     1141 atggtgagca agggcgagga gaccacaatg ggcgtaatca agcccgacat gaagatcaag
     1201 ctgaagatgg agggcaacgt gaatggccac gccttcgtga tcgagggcga gggcgagggc
     1261 aagccctacg acggcaccaa caccatcaac ctggaggtga aggagggagc ccccctgccc
     1321 ttctcctacg acattctgac caccgcgttc gcccttggca acagggcctt caccaagtac
     1381 cccgacgaca tccccaacta cttcaagcag tccttccccg agggctactc ttgggagcgc
     1441 accatgacct tcgaggacaa gggcatcgtg aaggtgaagt ccgacatctc catggaggag
     1501 gactccttca tctacgagat acacctcaag ggcgagaact tcccccccaa cggccccgtg
     1561 atgcagaaga agaccaccgg ctgggacgcc tccaccgaga ggatgtacgt gcgcgacggc
     1621 gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg gcggcggcca ccaccgcgtt
     1681 gacttcaaga ccatctacag ggccaagaag gcggtgaagc tgcccgacta tcactttgtg
     1741 gaccaccgca tcgagatcct gaaccacgac aaggactaca acaaggtgac cgtttacgag
     1801 agcgccgtgg cccgcatgat gaagcggctg gtgtacatca gcaagatcag cggccacctg
     1861 agcctggaag agatccagcg gatcggcaag gtgtccatca agaacaacca gcgggacaac
     1921 atcaccggcg tgctgctgta cctgcagggc ctgttcttcc agatcctgga aggcgagaac
     1981 gagaaggtgg acaagctgta caagaaaatc ctggtggacg accggcacac caacatcctg
     2041 tgcctgaaaa ccgagtacga catcaccgac cggatgttcc ccaactgggc catgaaaacc
     2101 atcaacctga acgagaacag cgagctgatg atccagccca tcaagagcct gctgcagacc
     2161 atcacccaga gccaccgggt gctggaaaag tacatgcccg ccagagtgat ctacctgatc
     2221 aaccagggca tcaaccccct gaccgtggag ccccagctgg tcgagaagat catcttcttc
     2281 agcgacatcc tggccttcag caccctgacc gagaagctgc ccgtgaacga ggtggtcatc
     2341 ctggtcaacc ggtacttcag catctgcacc cggatcatca gcgcctacgg cggcgaagtg
     2401 accaagttca tcggcgactg cgtgatggcc agcttcacca aagagcaggg cgacgccgcc
     2461 atccggacca gcctggacat catctccgag ctgaagcagc tgcggcacca cgtggaggcc
     2521 accaaccccc tgcacctgct gtacaccggc atcggcctga gctacggcca cgtgatcgag
     2581 ggcaacatgg gcagcagcct gaagatggac cacaccctgc tgggggacgc cgtgaacgtg
     2641 gccgccaggc tggaagccct gacaagacag ctgccctacg ccctggcctt tacagccggc
     2701 gtgaagaagt gctgccaggc ccagtggacc ttcatcaacc tgggagccca ccaggtcaag
     2761 ggcaagcagg aagccatcga ggtctacacc gtgaatgagg cccagaagta ctacgacacc
     2821 ctgcagatca cccagctgat ccggcagacc ctggaaaacg acaaggcgac gaattttagt
     2881 ctactgaaac aagcgggaga cgtggaggaa aaccctggac ctatggtgag caagggcgag
     2941 gagaccacaa tgggcgtaat caagcccgac atgaagatca agctgaagat ggagggcaac
     3001 gtgaatggcc acgccttcgt gatcgagggc gagggcgagg gcaagcccta cgacggcacc
     3061 aacaccatca acctggaggt gaaggaggga gcccccctgc ccttctccta cgacattctg
     3121 accaccgcgt tcgcctacgg caacagggcc ttcaccaagt accccgacga catccccaac
     3181 tacttcaagc agtccttccc cgagggctac tcttgggagc gcaccatgac cttcgaggac
     3241 aagggcatcg tgaaggtgaa gtccgacatc tccatggagg aggactcctt catctacgag
     3301 atacacctca agggcgagaa cttccccccc aacggccccg tgatgcagaa gaagaccacc
     3361 ggctgggacg cctccaccga gaggatgtac gtgcgcgacg gcgtgctgaa gggcgacgtc
     3421 aagcacaagc tgctgctgga gggcggcggc caccaccgcg ttgacttcaa gaccatctac
     3481 agggccaaga aggcggtgaa gctgcccgac tatcactttg tggaccaccg catcgagatc
     3541 ctgaaccacg acaaggacta caacaaggtg accgtttacg agagcgccgt ggcccgcaac
     3601 tccaccgacg gcatggacga gctgtacaag taactcgaga gatctacggg tggcatccct
     3661 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     3721 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     3781 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     3841 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     3901 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     3961 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     4021 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     4081 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa
     4141 ccacgtgcgg accgaggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4201 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca
     4261 gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca gcaaaaggcc
     4321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4561 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4621 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4681 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4741 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4801 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4861 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4921 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4981 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5041 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5101 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5161 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5221 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5281 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5341 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5401 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5461 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5521 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5581 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5641 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5701 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5761 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5821 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5881 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5941 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6001 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6061 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6121 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6181 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     6241 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     6301 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataaa
     6361 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     6421 tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata
     6481 gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     6541 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa
     6601 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     6661 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     6721 aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     6781 cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga cgtatgcggt
     6841 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc tgcaggcagc
     6901 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
     6961 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     7021 ggggttcctg cggccaagcg tggagctagt tattaatagt aatcaattac ggggtcatta
     7081 gttc
//
