LOCUS       Exported                7686 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gq-coupled hM3D-IRES-mCitrine under the control of human synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-HA-hM3D(Gq)-IRES-mCitrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7686)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7686)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7686
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             674..700
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2486..3059
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2653..2670)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2880..2899
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3060..3069
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3066..3782
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3111..3132)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3372..3391)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3719..3740
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3813..4401
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3866..3886)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4284..4295)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4433..4909
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4632..4651)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4949..5089
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4949..5078
                     /label=AAV2 ITR
     rep_origin      5164..5619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5251..5270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5461..5482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5636..5655)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5755..5777
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5815..5833)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5901..6005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6006..6866
                     /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(6224..6243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7037..7625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7526..7545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgacgccacc atgtacccat acgatgttcc agattacgct accttgcaca ataacagtac
      721 aacctcgcct ttgtttccaa acatcagctc ctcctggata cacagcccct ccgatgcagg
      781 gctgcccccg ggaaccgtca ctcatttcgg cagctacaat gtttctcgag cagctggcaa
      841 tttctcctct ccagacggta ccaccgatga ccctctggga ggtcataccg tctggcaagt
      901 ggtcttcatc gctttcttaa cgggcatcct ggccttggtg accatcatcg gcaacatcct
      961 ggtaattgtg tcatttaagg tcaacaagca gctgaagacg gtcaacaact acttcctctt
     1021 aagcctggcc tgtgccgatc tgattatcgg ggtcatttca atgaatctgt ttacgaccta
     1081 catcatcatg aatcgatggg ccttagggaa cttggcctgt gacctctggc ttgccattga
     1141 ctgcgtagcc agcaatgcct ctgttatgaa tcttctggtc atcagctttg acagatactt
     1201 ttccatcacg aggccgctca cgtaccgagc caaacgaaca acaaagagag ccggtgtgat
     1261 gatcggtctg gcttgggtca tctcctttgt cctttgggct cctgccatct tgttctggca
     1321 atactttgtt ggaaagagaa ctgtgcctcc gggagagtgc ttcattcagt tcctcagtga
     1381 gcccaccatt acttttggca cagccatcgc tggtttttat atgcctgtca ccattatgac
     1441 tattttatac tggaggatct ataaggaaac tgaaaagcgt accaaagagc ttgctggcct
     1501 gcaagcctct gggacagagg cagagacaga aaactttgtc caccccacgg gcagttctcg
     1561 aagctgcagc agttacgaac ttcaacagca aagcatgaaa cgctccaaca ggaggaagta
     1621 tggccgctgc cacttctggt tcacaaccaa gagctggaaa cccagctccg agcagatgga
     1681 ccaagaccac agcagcagtg acagttggaa caacaatgat gctgctgcct ccctggagaa
     1741 ctccgcctcc tccgacgagg aggacattgg ctccgagacg agagccatct actccatcgt
     1801 gctcaagctt ccgggtcaca gcaccatcct caactccacc aagttaccct catcggacaa
     1861 cctgcaggtg cctgaggagg agctggggat ggtggacttg gagaggaaag ccgacaagct
     1921 gcaggcccag aagagcgtgg acgatggagg cagttttcca aaaagcttct ccaagcttcc
     1981 catccagcta gagtcagccg tggacacagc taagacttct gacgtcaact cctcagtggg
     2041 taagagcacg gccactctac ctctgtcctt caaggaagcc actctggcca agaggtttgc
     2101 tctgaagacc agaagtcaga tcactaagcg gaaaaggatg tccctggtca aggagaagaa
     2161 agcggcccag accctcagtg cgatcttgct tgccttcatc atcacttgga ccccatacaa
     2221 catcatggtt ctggtgaaca ccttttgtga cagctgcata cccaaaacct tttggaatct
     2281 gggctactgg ctgtgctaca tcaacagcac cgtgaacccc gtgtgctatg ctctgtgcaa
     2341 caaaacattc agaaccactt tcaagatgct gctgctgtgc cagtgtgaca aaaaaaagag
     2401 gcgcaagcag cagtaccagc agagacagtc ggtcattttt cacaagcgcg cacccgagca
     2461 ggccttgtag gcggccgcga tatcgcccct ctccctcccc cccccctaac gttactggcc
     2521 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2581 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2641 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2701 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2761 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2821 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2881 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2941 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     3001 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
     3061 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3121 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3181 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3241 ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca
     3301 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3361 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3421 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3481 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3541 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3601 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3661 accactacct gagctaccag tccaaactga gcaaagaccc caacgagaag cgcgatcaca
     3721 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3781 agtgatagga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     3841 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3901 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3961 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4021 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4081 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4141 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4201 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     4261 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4321 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     4381 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     4441 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     4501 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4561 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4621 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4681 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4741 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4801 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4861 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     4921 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4981 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     5041 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5101 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5161 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5221 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5281 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5341 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5401 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5461 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     5521 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5581 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5641 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5701 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5761 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5821 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5881 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5941 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6001 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6061 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6121 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6181 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6241 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6301 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6361 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6421 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6481 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6541 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6601 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6661 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6721 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6781 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6841 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6901 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6961 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7021 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7081 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7141 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7201 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7261 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7321 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7381 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7441 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7501 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7561 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7621 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7681 acatgt
//
