LOCUS       Exported                7667 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses trkB.T1 fused with mCherry in a cre dependent manner..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-DIO-trkB.DN-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7667)
  AUTHORS   Guyon N, Zacharias LR, van Lunteren JA, Immenschuh J, Fuzik J, 
            Martin A, Xuan Y, Zilberter M, Kim H, Meletis K, Lopes-Aguiar C, 
            Carlen M
  TITLE     Adult trkB signaling in parvalbumin interneurons is essential to 
            prefrontal network dynamics.
  JOURNAL   J Neurosci. 2021 Feb 15. pii: JNEUROSCI.1848-20.2021. doi: 
            10.1523/JNEUROSCI.1848-20.2021.
  PUBMED    33593856
REFERENCE   2  (bases 1 to 7667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7667
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   36..165
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        266..1444
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          497..1435
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1392..1412
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1475..1508
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    1559..1592
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1601..2308)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1879..1898)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2148..2166
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    complement(3687..3720)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(3771..3804)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3829..4417
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3882..3902)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4300..4311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4449..4925
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4648..4667)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4965..5105
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4965..5094
                     /label=AAV2 ITR
     rep_origin      5180..5635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5267..5286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5477..5498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5652..5671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5771..5793
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5831..5849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5917..6021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6022..6882
                     /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(6240..6259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7053..7641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7542..7561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
       61 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
      121 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgta
      181 agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt
      241 ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc
      301 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      361 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      421 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      481 tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg
      541 ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc
      601 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      661 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      721 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      781 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      841 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      901 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      961 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
     1021 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
     1081 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
     1141 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1201 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1261 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1321 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1381 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1441 gtgaggtacc ggatcctcta gagtcgactc cggaataact tcgtatagga tactttatac
     1501 gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat
     1561 aacttcgtat agcatacatt atacgaagtt atggcgcgcc ttacttgtac agctcgtcca
     1621 tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct
     1681 cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg
     1741 gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct
     1801 tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg
     1861 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc
     1921 cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg
     1981 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     2041 acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca
     2101 tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca
     2161 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     2221 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
     2281 ccatgttatc ctcctcgccc ttgctcactg ctgctgcgag cagaagcagc atcaccagca
     2341 ggcagaatcc taccacagag gcaatcacca ccacagcata gaccgagaga tgctcccgat
     2401 tggtttggtc agcaacatcc gtggagggga tctcattact tttgtttgta gtatccccga
     2461 tgtcagttgg cgtggtccag tcttcataga ggacttcagg gtaatttggg tttgtctcat
     2521 agtcaactcc aggccggccc atgaagtgag cagaaatctg tctctcgtcc ttcccatatt
     2581 cattcttggc cattagggtg tagtctccat tattcatatg agtggggtta tccagctgga
     2641 ggcagccgtg gtactccgtg tgattggtga cgtgtatttt ggtacagatg tacttggatt
     2701 cattcagtat ggctccgttg tagaaccact gaagtgctgg cttggggttg cctctcacag
     2761 tgaatgggat gcaccagtgg tggtctgagg ttggagattc gagaaatgtg atggttggtg
     2821 caaaatgcac agtgaggttc acagagtctt gatcttctcc gacgaggttt tctgccacac
     2881 aagagatttg tttcccactg tcatccgatg aaatgtttgt tatccttaag gagccctgtg
     2941 tgtggcttgt ttcattcatg tgtttggaaa ccaaattccc aacgtcccag tacaaggtgg
     3001 ggagcgggtc acccccgacg ctgcaggaaa tggtcacaga cttcccttcc tccaccgtga
     3061 ggttaggagc ggccagacgt gcagacggca gaccacaatt gggaatctgc aggttcgcca
     3121 gaggggtatt cttgctgctc tcattgaggc aatacaaatc ctgagtgtcg gggctggatt
     3181 tcgtctcctg gagagtcttg agccacatga tgtcacagga acacgtgaac ggattacccg
     3241 tcaggatcag gtcagacaag tcaaggtggc ggaaatgtct cctggacaaa ctcgtcagct
     3301 tgtttcgagt gaaattgatg tgccgcaggt tgccgttctt cagaaacgcc ttgtaagcca
     3361 caaactttaa gccggaatcc acaattgtaa ggtttttcag ccccacgtaa gcttcgacat
     3421 catcttcatt gatgatttct aaccttttct ggtttgcaat gagaatttcg gtgatgttct
     3481 ctgggtcaat gctgttaggt tccaacctcg gaaatgccac gatgccagga gaaggctcgg
     3541 tacaccaaat cctagtggtg ctgcatttgc aggacatggg gcaggcaaga gaagccctcc
     3601 agaagcccaa gaccagcaag cataagcccc agagccgcgc catggcgggt ccatgccacc
     3661 tcggccaggg cgacatggtg gctagcataa cttcgtataa agtatcctat acgaagttat
     3721 ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt
     3781 ataatgtatg ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg
     3841 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     3901 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     3961 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4021 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4081 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4141 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4201 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     4261 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4321 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4381 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag
     4441 agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     4501 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     4561 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     4621 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     4681 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     4741 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     4801 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     4861 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     4921 tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat
     4981 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     5041 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     5101 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5161 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5221 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     5281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     5401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     5521 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     5581 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     5641 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     5701 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     5761 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     5821 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5881 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6001 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6061 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6121 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6181 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6241 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6301 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6361 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6421 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6481 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6541 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6601 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6661 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6721 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6781 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6841 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6901 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6961 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacg
//
