LOCUS       Exported                7163 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV construct with tAKAR-alpha being expressed under the human 
            synapsin promotor with WPRE enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSYNCMV tAKAR-alpha
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  AUTHORS   Ma L, Jongbloets BC, Xiong WH, Melander JB, Qin M, Lameyer TJ, 
            Harrison MF, Zemelman BV, Mao T, Zhong H
  TITLE     A Highly Sensitive A-Kinase Activity Reporter for Imaging 
            Neuromodulatory Events in Awake Mice.
  JOURNAL   Neuron. 2018 Aug 22;99(4):665-679.e5. doi: 
            10.1016/j.neuron.2018.07.020. Epub 2018 Aug 9.
  PUBMED    30100256
REFERENCE   2  (bases 1 to 7163)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7163
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        71..373
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        415..862
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      873..882
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             879..1397
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     1532..1553
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3462..4050
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3515..3535)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3933..3944)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4075..4196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4086..4105)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4140..4159
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4296..4425
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4487..5075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4567..4586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5246..6106)
                     /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     5869..5888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6107..6211)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6279..6297
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6335..6357)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6457..6476
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6493..6948)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6630..6651)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6842..6861
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7023..7163
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7034..7163
                     /label=AAV2 ITR
ORIGIN
        1 gcggccgcgg tacctattaa tagtaatcaa ttacggggtc attagttcat agcccatata
       61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      361 tcgctattac cattcagaac cacggacagc accgaattca gggccctgcg tatgagtgca
      421 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      481 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      541 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      601 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      661 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      721 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      781 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      841 cgtgtcgtgc ctgagagcgc agtcggccgg tcgccaccat ggtgagcaag ggcgaggagc
      901 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      961 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1021 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1081 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1141 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1201 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1261 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1321 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1381 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1441 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     1501 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1561 cccgcatgca taagttttct caagaacaga tcggcgaaaa cattgtgtgc agggtcattt
     1621 gtaccacggg tcaaattccc atccgagatt tgtcagctga tatttcacaa gtgcttaagg
     1681 aaaaacgatc cataaagaaa gtttggacat ttggtagaaa cccagcctgt gactatcatt
     1741 taggaaacat ttcaagactg tcaaataagc atttccaaat actactagga gaagacggta
     1801 accttttatt gaatgacatt tccactaatg ggacctggtt aaatgggcaa aaagtcgaga
     1861 agaacagcaa tcagttactg tctcaaggtg atgaaataac cgttggtgta ggcgtggaat
     1921 cagatatttt atctctggtc attttcataa acgacaaatt taagcagtgc ctcgagcaga
     1981 acaaagttga tcgctctgca ggtaagccag gcagcggcga gggcagcacc aagggcctgc
     2041 gtcgcgccac cctggtggac ggcggcaccg gcggcagcga gctcatgggc agcgtgcagc
     2101 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2161 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     2221 tggtcctgct ggagagagtg accgccgccg ggatcactct cggcatggac gagctataca
     2281 agggtggaac tggaggttca atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2341 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2401 gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc accggcaagc
     2461 tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc
     2521 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2581 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2641 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2701 acggcaacat cctggggcac aagctggagt acaactggaa cagcgtcaac gtctatatca
     2761 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagt
     2821 ccggagtgta caagggtacc ccaggaaccc ccaagtctgc catcttggtg ccgagtgaga
     2881 agaaggtcgc catcatacgt actcctccaa aatctcctgc gactcccaag cagcttcggc
     2941 ttattaacca accactgcca gacctgaaga atgtcaaatc caaaatcgga tcaacagaca
     3001 acatcaaata ccagcctaaa ggggggcagg tacaaattgt taccaagaaa atagacctaa
     3061 gccatgtgac atccaaatgt ggctctctga agaacatccg ccacaggcca ggtggcggac
     3121 gtgtgaaaat tgagagtgta aaactagatt tcaaagaaaa ggcccaagct aaagttggtt
     3181 ctcttgataa tgctcatcat gtacctggag gtggtaatgt caagattgac agccaaaagt
     3241 tgaacttcag agagcatgct aaagcccgtg tggaccatgg ggctgagatc attacacagt
     3301 ccccaggcag atccagcgtg gcatcacccc gacgactcag caatgtctcc tcgtctggaa
     3361 gcatcaacct gctcgaatct cctcagcttg ccactttggc tgaggatgtc actgctgcac
     3421 tcgctaagca gggcttgtaa tctagaacgg ccgttatcga taatcaacct ctggattaca
     3481 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3541 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3601 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3661 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3721 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3781 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3841 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     3901 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3961 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4021 gtcggatctc cctttgggcc gcctccccgc atcgattgcg gccgtcagac atgataagat
     4081 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4141 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4201 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4261 gcaagtaaaa cctctacaaa tgtggtagcg gccgcaggaa cccctagtga tggagttggc
     4321 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4381 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggacatg tgagcaaaag
     4441 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4501 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4561 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4621 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4681 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4741 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4801 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4861 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4921 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4981 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5041 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5101 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5161 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5221 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5281 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5341 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5401 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5461 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5521 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5581 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5641 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5701 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5761 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5821 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5881 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5941 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6001 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6061 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6121 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6181 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6241 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6301 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6361 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6421 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6481 gagtgcacca taaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt
     6541 aaatcagctc attttttaac caatagaccg aaatcggcaa aatcccttat aaatcaaaag
     6601 aatagcccga gatagagttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6661 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6721 aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6781 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6841 aagggaagaa agcgaaagga gcgggcgcta aggcgctggc aagtgtagcg gtcacgctgc
     6901 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtac tatggttgct
     6961 ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     7021 cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg
     7081 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     7141 caactccatc actaggggtt cct
//
