LOCUS       Exported                7403 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  To express hM3Dq in cells not expressing Cre..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV hSyn FLEx-OFF hM3Dq-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  AUTHORS   Beier KT, Kim CK, Hoerbelt P, Hung LW, Heifets BD, DeLoach KE, Mosca
            TJ, Neuner S, Deisseroth K, Luo L, Malenka RC
  TITLE     Rabies screen reveals GPe control of cocaine-triggered plasticity.
  JOURNAL   Nature. 2017 Sep 21;549(7672):345-350. doi: 10.1038/nature23888. 
            Epub 2017 Sep 13.
  PUBMED    28902833
REFERENCE   2  (bases 1 to 7403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7403
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      1826..1835
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1832..2542
                     /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(1977..1995)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2245..2264
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2552..2585
                     /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    2656..2689
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2737..3325
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2790..2810)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3208..3219)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3357..3833
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3556..3575)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3873..4013
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3873..4002
                     /label=AAV2 ITR
     rep_origin      4088..4543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4175..4194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4385..4406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4560..4579)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4679..4701
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4739..4757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4825..4929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4930..5790
                     /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(5148..5167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5961..6549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6450..6469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6611..6740
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        6785..7232
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(7308..7341)
                     /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)."
ORIGIN
        1 ataacttcgt ataatgtatg ctatacgaag ttatgtcgac atgaccttgc acaataacag
       61 tacaacctcg cctttgtttc caaacatcag ctcctcctgg atacacagcc cctccgatgc
      121 agggctgccc ccgggaaccg tcactcattt cggcagctac aatgtttctc gagcagctgg
      181 caatttctcc tctccagacg gtaccaccga tgaccctctg ggaggtcata ccgtctggca
      241 agtggtcttc atcgctttct taacgggcat cctggccttg gtgaccatca tcggcaacat
      301 cctggtaatt gtgtcattta aggtcaacaa gcagctgaag acggtcaaca actacttcct
      361 cttaagcctg gcctgtgccg atctgattat cggggtcatt tcaatgaatc tgtttacgac
      421 ctacatcatc atgaatcgat gggccttagg gaacttggcc tgtgacctct ggcttgccat
      481 tgactgcgta gccagcaatg cctctgttat gaatcttctg gtcatcagct ttgacagata
      541 cttttccatc acgaggccgc tcacgtaccg agccaaacga acaacaaaga gagccggtgt
      601 gatgatcggt ctggcttggg tcatctcctt tgtcctttgg gctcctgcca tcttgttctg
      661 gcaatacttt gttggaaaga gaactgtgcc tccgggagag tgcttcattc agttcctcag
      721 tgagcccacc attacttttg gcacagccat cgctggtttt tatatgcctg tcaccattat
      781 gactatttta tactggagga tctataagga aactgaaaag cgtaccaaag agcttgctgg
      841 cctgcaagcc tctgggacag aggcagagac agaaaacttt gtccacccca cgggcagttc
      901 tcgaagctgc agcagttacg aacttcaaca gcaaagcatg aaacgctcca acaggaggaa
      961 gtatggccgc tgccacttct ggttcacaac caagagctgg aaacccagct ccgagcagat
     1021 ggaccaagac cacagcagca gtgacagttg gaacaacaat gatgctgctg cctccctgga
     1081 gaactccgcc tcctccgacg aggaggacat tggctccgag acgagagcca tctactccat
     1141 cgtgctcaag cttccgggtc acagcaccat cctcaactcc accaagttac cctcatcgga
     1201 caacctgcag gtgcctgagg aggagctggg gatggtggac ttggagagga aagccgacaa
     1261 gctgcaggcc cagaagagcg tggacgatgg aggcagtttt ccaaaaagct tctccaagct
     1321 tcccatccag ctagagtcag ccgtggacac agctaagact tctgacgtca actcctcagt
     1381 gggtaagagc acggccactc tacctctgtc cttcaaggaa gccactctgg ccaagaggtt
     1441 tgctctgaag accagaagtc agatcactaa gcggaaaagg atgtccctgg tcaaggagaa
     1501 gaaagcggcc cagaccctca gtgcgatctt gcttgccttc atcatcactt ggaccccata
     1561 caacatcatg gttctggtga acaccttttg tgacagctgc atacccaaaa ccttttggaa
     1621 tctgggctac tggctgtgct acatcaacag caccgtgaac cccgtgtgct atgctctgtg
     1681 caacaaaaca ttcagaacca ctttcaagat gctgctgctg tgccagtgtg acaaaaaaaa
     1741 gaggcgcaag cagcagtacc agcagagaca gtcggtcatt tttcacaagc gcgcacccga
     1801 gcaggccttg aaggatcccc cggtcgccac catggtgagc aagggcgagg aggataacat
     1861 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1921 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1981 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2041 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2101 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2161 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2221 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2281 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2341 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2401 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2461 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2521 catggacgag ctgtacaagt aaggcgcgcc tataacttcg tataggatac tttatacgaa
     2581 gttatcattg ggattcttcc tattttgatc caagcatcac catcgaccct ctagtccaga
     2641 tctcaccatc gacccataac ttcgtatagc atacattata cgaagttata agcttatatg
     2701 cggccgatcg atgaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     2761 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2821 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2881 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2941 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3001 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3061 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3121 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3181 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3241 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3301 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     3361 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     3421 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     3481 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     3541 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     3601 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     3661 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     3721 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     3781 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     3841 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3901 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3961 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     4021 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4081 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4141 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4201 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4261 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4321 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4381 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4441 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4501 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4561 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4621 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4681 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4741 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4861 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4921 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4981 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5041 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5101 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5161 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5221 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5281 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5341 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5401 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5461 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5521 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5581 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5641 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5701 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5761 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5821 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6601 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     6661 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     6721 ctccatcact aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg
     6781 tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga
     6841 ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat
     6901 cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac
     6961 cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag
     7021 gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg
     7081 tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg
     7141 ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca
     7201 gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctgtgctgtc
     7261 tcatcatttt ggcaaagaat tgatttgata ccgcgggccc ggccgggata acttcgtata
     7321 aagtatccta tacgaagtta tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa
     7381 ttgccaagca tcaccatcga cac
//
