LOCUS       Exported                8127 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV vector to drive the expression of dTomato in PV cortical 
            interneuronsunder the control of the E7 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E7-dTom-nls-dTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  AUTHORS   Vormstein-Schneider DC, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias Garcia M, Sakopoulos S, Stevenson O, Schneider G, Zhang Q, 
            Sharma J, Franken TP, Smith J, Vogel I, Sanchez V, Ibrahim LA, 
            Burbridge T, Favuzzi E, Saldi GA, Xu Q, Guo L, Yuan X, Zaghloul KA, 
            Sabri E, Goldberg EM, Devinsky O, Batista-Brito R, Reynolds J, Feng 
            G, Fu Z, McBain CJ, Fishell GJ, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans
  JOURNAL   Nature Neuroscience (2020)
REFERENCE   2  (bases 1 to 8127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8127
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          2746..2878
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      2901..2910
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2907..3608
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     CDS             3630..3686
                     /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             3699..3719
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3720..4418
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     misc_feature    4473..5061
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4526..4546)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4944..4955)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5230..5311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5267..5286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5321..5340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5390..5530
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5390..5519
                     /label=AAV2 ITR
     rep_origin      5605..6060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5902..5923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6077..6096)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6196..6218
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6256..6274)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6342..6446
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6447..7307
                     /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(6665..6684)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7478..8066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7967..7986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaatt aaactagtgg tttgatactg tataattaat
      181 taggcctcca atcatgcctt cccagcctcc acggatggag aaaccctctc cgccatgcct
      241 taaagaggaa ttgctgtaat aaatgagtct cctgatagca aatttctcag caagggggaa
      301 tcgcgtaaat ggagacatag tattgacagc aaagtccaat gtgttatttt taccagaacg
      361 aactctccgg ttcaagcctt tgaaagagac atttgaaaac caaaaacaaa caatgtaatg
      421 gagcgaggaa aaaagccaca gaagtgagtg gcagggagtt taaaagagca gatgccactg
      481 ccaggtctat gggacataac cagccacttg tgctgggtct tggcagttta taatgctacc
      541 tcatcttctc cgcgaaattg ttttcccgta aatctctgtg gccatccatt cctgtctaca
      601 cattatgttc ctaaaataga caccatctaa aaatcacttc aaggagcttt gtggaggaag
      661 gcctaaattg caacactcct ccagcgaaga tagatgcagt gtttgatggc attaccagtc
      721 ggtagccagg aaggggagtt tgtgaggagt ttttccacca cagttaatct gtttctggaa
      781 ggaaagggaa gtgtcagact tcccgaggag gcaaacgtgt gtggaagctc tcatttgcat
      841 cacccccggc ctgtcaggta ttgcagcaaa agggagaggt gagctaccct ggctctcctt
      901 gggcaggagg gacagaatca ggaagcatca acctcagcat ggaattttcc tattcctgtt
      961 tggcatcctc ctcttgggat gatttacagc gcgggttgga gaaacacgct ctgccactcc
     1021 actagcgcac cagatagaca gtgcagacct gcagatccat acccgaggag aagccacatt
     1081 tcctacgtgt gatagcaaca gcgtttggca atttgcgact ttgctactgc agcttagaaa
     1141 atatttagtc acatgcacat ctgaacagaa agacacccag gcttgactca gtcatttccg
     1201 tcagacacac gaaagaaaaa gcgtctctgc tcacaagctt atttggactg ctttgttgaa
     1261 aggaggggcg gcagacactt tgtagatgtg gcaagagggc tttatatcca gacctcaaac
     1321 aggtaggaga gaaggaagcc aggagaggta aggaaggggc gtggaaaagc ctcacagcca
     1381 cctcgaagaa aacagttttt ttgccctgtt cagaaagcaa gaggttccac agtggttttg
     1441 tgtcaatgga gcacatctgc agtatcattg ccgttggtga cctctgtcta attaaaagta
     1501 agtcagtcct tcccacccgg cattgtctga aacccgggac tctttatcac tttgctaaag
     1561 ttcatttgca agtgtagtta aggaagagtc aggggggaaa cagcatctgt cccttctggt
     1621 cctggggagg aggcactcct ttccaagagt caagcctctg cccaaagaag ctgcctcccc
     1681 tgcaatgcta ggatccagga gcagccccgc tgccttcttg cttcctctgt gaggtctaat
     1741 ttttgcatca tctttaggag cgatatgacc tctattcaca gccatcgaat ccagttccaa
     1801 agcaccaatg acagaggggg cttcaagaca agaccttgcc taggaggatg caggcaagca
     1861 aaggcaagag ctggcccgat gccaagttat tttaggccaa agaatctcat ccttctatca
     1921 aaatgctgaa ctgcaaaacg aacctgattt cagttcatgg aaggttgaga ggaggaggag
     1981 ggggagggga ggaggaggaa gagaggagga ggggaggagg agggaggagg aggaggggag
     2041 gaggaggggg acagttggtc cgaattcaca tgcaaaaata gacttcctgt tctgccccaa
     2101 ctcttatttc cgtgggctct tctccccaag gatttaccag gtaagaattc accaccaaag
     2161 aagatcacaa tgagataatc agatggctta cctgataaaa aggaaaatta tccatctgca
     2221 gtgaggagca acatctcccc acgacgagtc cgcaccttcc gttgcaacga ttcagattcc
     2281 ttcttgcaaa aggtgaccaa gtgcttcaca agggctgcag cctcataggg gcagaacaca
     2341 cgtacacaaa cacacgcaca cacacacaca catgcaccag agacctctgc agtatcctct
     2401 cggcttcatc ctcgcctcac tctatggtac ctaatacaaa tcagcaaata gcttgtttta
     2461 aaaaaaaaag aaagaaaaaa aagcggagac agcacctaac gttacagtgc catctagtgg
     2521 ctacatcgta aataggttct cacagcctgg atttctgtgt tctttctcaa ccgcttcctt
     2581 ctggttcctt tttaaacttc ctgcagcccg ggctgggcat aaaagtcagg gcagagccat
     2641 ctattgctta catttgcttc tagcctgcaa gtcgaggagc gcagccttcc agaagcagag
     2701 cgcggcgcct taagctgcag aagttggtcg tgaggcactg ggcacgtaag tatcaaggtt
     2761 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     2821 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacagcg
     2881 agctcgctag cgcgatcgct gccaccatgg tgagcaaggg cgaggaggtc atcaaagagt
     2941 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     3001 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3061 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     3121 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     3181 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     3241 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     3301 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     3361 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     3421 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     3481 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     3541 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc
     3601 tgtacaagtc cggagataac ggaagcggag ccactaactt ctccctgttg aaacaagcag
     3661 gggatgtcga agagaatccc gggccatcta gaatggcccc caagaagaag aggaaggtgg
     3721 tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct
     3781 ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3841 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
     3901 tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg
     3961 attacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     4021 acggcggtct ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca
     4081 aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca
     4141 tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga
     4201 tccaccaggc cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct
     4261 acatggccaa gaagcccgtg caactgcccg gctactacta cgtggacacc aagctggaca
     4321 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc
     4381 accacctgtt cctgtacggc atggacgagc tgtacaagta agtcgacggc gcgccccacc
     4441 cctgcaggga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     4501 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     4561 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4621 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4681 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4741 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4801 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4861 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     4921 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4981 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     5041 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct gtgatagcgg
     5101 ccatcaagct ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     5161 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     5221 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5341 aatgtatcag cttatcgata ccgcatgcac gtgcggaccg agcggccgca ggaaccccta
     5401 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     5461 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     5521 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5581 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     5701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5761 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5821 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     5941 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     6001 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     6061 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6121 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6181 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6241 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6301 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6361 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6421 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6481 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6541 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6601 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6661 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6721 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6781 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6841 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6901 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6961 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7021 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7081 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7141 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7201 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7261 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7321 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7381 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7621 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8101 ccttttgctg gccttttgct cacatgt
//
