LOCUS       Exported                6726 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Gi-coupled hM4D DREADD fused with mCherry under the control of human
            synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-hM4D(Gi)-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  AUTHORS   Bryan Roth
  TITLE     Roth lab DREADDs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6726
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      654..663
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2112..2121
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2118..2828
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2263..2281)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2531..2550
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2853..3441
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2906..2926)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3324..3335)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3473..3949
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3672..3691)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3989..4129
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3989..4118
                     /label=AAV2 ITR
     rep_origin      4204..4659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4291..4310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4501..4522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4676..4695)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4795..4817
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4855..4873)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4941..5045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5046..5906
                     /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(5264..5283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6077..6665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6566..6585
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacgccacca
      661 tggccaactt cacacctgtc aatggcagct cgggcaatca gtccgtgcgc ctggtcacgt
      721 catcatccca caatcgctat gagacggtgg aaatggtctt cattgccaca gtgacaggct
      781 ccctgagcct ggtgactgtc gtgggcaaca tcctggtgat gctgtccatc aaggtcaaca
      841 ggcagctgca gacagtcaac aactacttcc tcttcagcct ggcgtgtgct gatctcatca
      901 taggcgcctt ctccatgaac ctctacaccg tgtacatcat caagggctac tggcccctgg
      961 gcgccgtggt ctgcgacctg tggctggccc tggactgcgt ggtgagcaac gcctccgtca
     1021 tgaaccttct catcatcagc tttgaccgct acttctgcgt caccaagcct ctcacctacc
     1081 ctgcccggcg caccaccaag atggcaggcc tcatgattgc tgctgcctgg gtactgtcct
     1141 tcgtgctctg ggcgcctgcc atcttgttct ggcagtttgt ggtgggtaag cggacggtgc
     1201 ccgacaacca gtgcttcatc cagttcctgt ccaacccagc agtgaccttt ggcacagcca
     1261 ttgctggctt ctacctgcct gtggtcatca tgacggtgct gtacatccac atctccctgg
     1321 ccagtcgcag ccgagtccac aagcaccggc ccgagggccc gaaggagaag aaagccaaga
     1381 cgctggcctt cctcaagagc ccactaatga agcagagcgt caagaagccc ccgcccgggg
     1441 aggccgcccg ggaggagctg cgcaatggca agctggagga ggcccccccg ccagcgctgc
     1501 caccgccacc gcgccccgtg gctgataagg acacttccaa tgagtccagc tcaggcagtg
     1561 ccacccagaa caccaaggaa cgcccagcca cagagctgtc caccacagag gccaccacgc
     1621 ccgccatgcc cgcccctccc ctgcagccgc gggccctcaa cccagcctcc agatggtcca
     1681 agatccagat tgtgacgaag cagacaggca atgagtgtgt gacagccatt gagattgtgc
     1741 ctgccacgcc ggctggcatg cgccctgcgg ccaacgtggc ccgcaagttc gccagcatcg
     1801 ctcgcaacca ggtgcgcaag aagcggcaga tggcggcccg ggagcgcaaa gtgacacgaa
     1861 cgatctttgc cattctgctg gccttcatcc tcacctggac gccctacaac gtcatggtcc
     1921 tggtgaacac cttctgccag agctgcatcc ctgacacggt gtggtccatt ggctactggc
     1981 tctgctacgt caacagcacc atcaaccctg cctgctatgc tctgtgcaac gccaccttta
     2041 aaaagacctt ccggcacctg ctgctgtgcc agtatcggaa catcggcact gccaggcggg
     2101 atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     2161 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     2221 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     2281 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     2341 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     2401 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     2461 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     2521 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     2581 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     2641 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     2701 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     2761 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     2821 acaagtaaga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     2881 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2941 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3001 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3061 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3121 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3181 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3241 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     3301 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3361 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3421 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     3481 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     3541 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     3601 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     3661 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     3721 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     3781 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3841 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3901 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     3961 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     4021 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     4081 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     4141 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     4201 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4261 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4321 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4381 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     4441 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4501 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     4561 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     4621 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     4681 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     4741 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4801 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     4861 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5581 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6721 acatgt
//
