LOCUS       Exported                6529 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 E3 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-S5E3-dTom-nlsdTom
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6529)
  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 6529)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6529
                     /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          1148..1280
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1303..1312
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1309..2010
                     /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             2032..2088
                     /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             2101..2121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2122..2820
                     /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    2875..3463
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2928..2948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3346..3357)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3632..3713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3669..3688)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3723..3742
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3792..3932
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3792..3921
                     /label=AAV2 ITR
     rep_origin      4007..4462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4094..4113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4304..4325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4479..4498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4598..4620
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4658..4676)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4744..4848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4849..5709
                     /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(5067..5086)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5880..6468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6369..6388
                     /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 tttataaaat tttattttcc
      181 taaaatttac atttaaaagc catccaactt accaaagtga tttcaaacca caattacatt
      241 ttatctcaac taccagcatt ttatttagcc aggatctaca tgagacacat atcatgatgt
      301 gctatgtaca tcttgttata cagtgttata ttgataacaa atgtcatcaa tataacattg
      361 aattaatctt ccataattta tggggaaaaa aggagcagcc ttactgaagg gcaaagttat
      421 acaacagctt tacagaagct gcatgcgagt gcagtaccgg gacacgggca cggacggcgg
      481 cactataacc attttccgtg gtggtaatct tgctttcatc tgacacagaa aagagaccgc
      541 cgttttgaaa actcacagaa ctagcctcac ggttttgtga gtccattgag cgctggctgc
      601 gaagaacggt gtttaactcg agaaatcatt gaacaagttt tagaaaataa agatgcttat
      661 gacaatttca aacttgaagg tctccaaaga aggactgaga tattggtgag aggagtaaag
      721 agaatcctgg tgcatttatt tcatgcttcc ttctgttcga agatcatttt gaggtttata
      781 aaaggtgggg tgatccaaaa atctccaggc tgagagtcct ggctgaggct gtgaactggg
      841 ctgcagagaa agggccacgc ctccctcctc tgctcgcatt actcagcagc ttttctgcat
      901 gtggctggct gcagacaatc taaacccttc cgctgtcgct ccccccttat actgttctgc
      961 caaaaggaag gcagagagga aatcagctac ggggcaaact tcctgcagcc cgggctgggc
     1021 ataaaagtca gggcagagcc atctattgct tacatttgct tctagcctgc aagtcgagga
     1081 gcgcagcctt ccagaagcag agcgcggcgc cttaagctgc agaagttggt cgtgaggcac
     1141 tgggcacgta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     1201 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     1261 ctttgccttt ctctccacag cgagctcgct agcgcgatcg ctgccaccat ggtgagcaag
     1321 ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac
     1381 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     1441 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     1501 cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag
     1561 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt
     1621 ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag
     1681 atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg
     1741 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag
     1801 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc
     1861 aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc
     1921 cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg
     1981 ttcctgtacg gcatggacga gctgtacaag tccggagata acggaagcgg agccactaac
     2041 ttctccctgt tgaaacaagc aggggatgtc gaagagaatc ccgggccatc tagaatggcc
     2101 cccaagaaga agaggaaggt ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
     2161 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     2221 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     2281 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     2341 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     2401 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     2461 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     2521 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     2581 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     2641 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     2701 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2761 tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag
     2821 taagtcgacg gcgcgcccca cccctgcagg gaattcgata tcaagcttat cgataatcaa
     2881 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2941 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3001 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3061 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3121 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3181 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3241 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     3301 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3361 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3421 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg
     3481 ctcgagagat ctgtgatagc ggccatcaag ctggccgcga ctctagatca taatcagcca
     3541 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3601 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3661 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3721 ttgtggtttg tccaaactca tcaatgtatc agcttatcga taccgcatgc acgtgcggac
     3781 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3841 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3901 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3961 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     4021 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4081 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4141 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4201 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     4261 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4321 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     4381 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4441 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     4501 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4561 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4621 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4681 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4741 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4801 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4861 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5701 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5761 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5821 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5881 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5941 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6001 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6061 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6121 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6181 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6241 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6301 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6361 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6421 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
