LOCUS       Exported                5742 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV-mediated expression of mEGFP and paAIP2 (R5AR6A), the functional
            mutant of paAIP2.
ACCESSION   .
VERSION     .
KEYWORDS    CaMKIIP-mEGFP-P2A-paAIP2 (R5AR6A)/pAAV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  AUTHORS   Murakoshi H, Shin ME, Parra-Bueno P, Szatmari EM, Shibata AC, Yasuda
            R
  TITLE     Kinetics of Endogenous CaMKII Required for Synaptic Plasticity 
            Revealed by Optogenetic Kinase Inhibitor.
  JOURNAL   Neuron. 2017 Apr 5;94(1):37-47.e5. doi: 
            10.1016/j.neuron.2017.02.036. Epub 2017 Mar 16.
  PUBMED    28318784
REFERENCE   2  (bases 1 to 5742)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5742
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   53..182
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        226..1514
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1537..1546
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1543..2259
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1588..1609)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1849..1868)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2196..2217
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2269..2325
                     /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"
     repeat_region   3057..3197
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3057..3186
                     /label=AAV2 ITR
     rep_origin      3272..3727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3359..3378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3569..3590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3744..3763)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3863..3885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3923..3941)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4009..4113
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4114..4974
                     /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     complement(4332..4351)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5145..5733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5634..5653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg
       61 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc
      121 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc
      181 ctgcggccgc acgcgtatta ccgccatgca ttagttatta attaacatta tggccttagg
      241 tcacttcatc tccatggggt tcttcttctg attttctaga aaatgagatg ggggtgcaga
      301 gagcttcctc agtgacctgc ccagggtcac atcagaaatg tcagagctag aacttgaact
      361 cagattacta atcttaaatt ccatgccttg ggggcatgca agtacgatat acagaaggag
      421 tgaactcatt agggcagatg accaatgagt ttaggaaaga agagtccagg gcagggtaca
      481 tctacaccac ccgcccagcc ctgggtgagt ccagccacgt tcacctcatt atagttgcct
      541 ctctccagtc ctaccttgac gggaagcaca agcagaaact gggacaggag ccccaggaga
      601 ccaaatcttc atggtccctc tgggaggatg ggtggggaga gctgtggcag aggcctcagg
      661 aggggccctg ctgctcagtg gtgacagata ggggtgagaa agcagacaga gtcattccgt
      721 cagcattctg ggtctgtttg gtacttcttc tcacgctaag gtggcggtgt gatatgcaca
      781 atggctaaaa agcagggaga gctggaaaga aacaaggaca gagacagagg ccaagtcaac
      841 cagaccaatt cccagaggaa gcaaagaaac cattacagag actacaaggg ggaagggaag
      901 gagagatgaa ttagcttccc ctgtaaacct tagaacccag ctgttgccag ggcaacgggg
      961 caatacctgt ctcttcagag gagatgaagt tgccagggta actacatcct gtctttctca
     1021 aggaccatcc cagaatgtgg cacccactag ccgttaccat agcaactgcc tctttgcccc
     1081 acttaatccc atcccgtctg ttaaaagggc cctatagttg gaggtggggg aggtaggaag
     1141 agcgatgatc acttgtggac taagtttgtt cgcatcccct tctccaaccc cctcagtaca
     1201 tcaccctggg ggaacagggt ccacttgctc ctgggcccac acagtcctgc agtattgtgt
     1261 atataaggcc agggcaaaga ggagcaggtt ttaaagtgaa aggcaggcag gtgttgggga
     1321 ggcagttacc ggggcaacgg gaacagggcg tttcggaggt ggttgccatg gggacctgga
     1381 tgctgacgaa ggctcgcgag gctgtgagca gccacagtgc cctgctcaga agccccaagc
     1441 tcgtcagtca agccggttct ccgtttgcac tcaggagcac gggcaggcga gtggccccta
     1501 gttctggggg cagcggggga tccccgggta ccggtcgcca ccatggtgag caagggcgag
     1561 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1621 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1681 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1741 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1801 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1861 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1921 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1981 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2041 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2101 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2161 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2221 gccgccggga tcactctcgg catggacgag ctgtacaagg gaagcggagc cactaacttc
     2281 tccctgttga aacaagcagg ggatgtcgaa gagaatcccg ggccaggatc catgctggcg
     2341 accaccctgg aacgcattga aaaaaacttt gtgattaccg atccgcgcct gccggataac
     2401 ccgattattt ttgcgagcga tagctttctg cagctgaccg aatatagccg cgaagaaatt
     2461 ctgggccgca actgccgctt tctgcagggc ccggaaaccg atcgcgcgac cgtgcgcaaa
     2521 attcgcgatg cgattgataa ccagaccgaa gtgaccgtgc agctgattaa ctataccaaa
     2581 agcggcaaaa aattttggaa cctgtttcat ctgcagccga tgcgcgatca gaaaggcgat
     2641 gtgcagtatt ttattggcgt gcagctggat ggcaccgaac atgtgcgcga tgcggcggaa
     2701 cgcgaaggcg tgatgctgat taaaaaaacc gcggaaaaca ttaagaagaa actggctgca
     2761 caggaagcgt ttgatgcgct gtaaaagctt gtggcatccc tgtgacccct ccccagtgcc
     2821 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     2881 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     2941 atggagcaag gggcaagttg ggaagaccca aattgctggg attacaggcg tgaaccactg
     3001 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     3061 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3121 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3181 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3241 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3301 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3361 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3421 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3481 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3541 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3601 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     3661 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     3721 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     3781 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     3841 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     3901 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     3961 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4021 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4081 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4141 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4201 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4261 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4321 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4381 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4441 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4501 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4561 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4621 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     4681 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     4741 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     4801 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     4861 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     4921 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     4981 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5041 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5101 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5161 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5221 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5281 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5341 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5401 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5461 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5521 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5581 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5641 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5701 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gc
//
