LOCUS       sequence-362037-        7808 bp    DNA     circular SYN 14-APR-2025
DEFINITION  INTRSECT Chemogenetics - expresses DREADD Gq-mCherry in cells 
            expressing Cre AND Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7808)
  TITLE     Deisseroth Lab Plasmids
REFERENCE   2  (bases 1 to 7808)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV-nEF Con/Fon DREADD Gq-mCherry
FEATURES             Location/Qualifiers
     source          1..7808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        177..388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             401..669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     primer_bind     complement(1104..1123)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(1269..1302)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1353..1386)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     regulatory      2360..2369
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2511..2529)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2728..2761
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2812..2845
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     misc_feature    3935..4523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3988..4008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4406..4417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4555..5031
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4754..4773)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5071..5211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5071..5200
                     /label=AAV2 ITR
     rep_origin      5286..5741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5373..5392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5583..5604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5758..5777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5877..5899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6023..6127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6128..6196,6197..6988)
                     /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(6346..6365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7159..7747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7648..7667
                     /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 gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
      421 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
      481 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
      541 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
      601 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
      661 ttacagatcc aagctgtgac cggcgcctac ggatccgaag ttcctattct tcaaatagta
      721 taggaacttc gcagagtagt agctggattg tagctgctat tagcaatcta aaacctctta
      781 gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg ccctattact tgtacagctc
      841 gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta
      901 gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg
      961 cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt
     1021 cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc
     1081 ggaggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg aggggaagtt
     1141 ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgtgg
     1201 gacgacatga cttaaccagt taaccatgtt ctctgaacat gagaagggct ggccactctc
     1261 cagtcgacat aacttcgtat aggatacttt atacgaagtt atgcagaatg gtagctggat
     1321 tgtagctgct attagcaata tgaaacctct taataacttc gtatagtata ccttatacga
     1381 agttatacta gttgtgcccc cccttttttt tataaaattg tattaatgtt attacatatc
     1441 tcctgtatgt gacccatgtg cttatgactc tatttctcat gtgtttagga ggaagtatgg
     1501 ccgctgccac ttctggttca caaccaagag ctggaaaccc agctccgagc agatggacca
     1561 agaccacagc agcagtgaca gttggaacaa caatgatgct gctgcctccc tggagaactc
     1621 cgcctcctcc gacgaggagg acattggctc cgagacgaga gccatctact ccatcgtgct
     1681 caagcttccg ggtcacagca ccatcctcaa ctccaccaag ttaccctcat cggacaacct
     1741 gcaggtgcct gaggaggagc tggggatggt ggacttggag aggaaagccg acaagctgca
     1801 ggcccagaag agcgtggacg atggaggcag ttttccaaaa agcttctcca agcttcccat
     1861 ccagctagag tcagccgtgg acacagctaa gacttctgac gtcaactcct cagtgggtaa
     1921 gagcacggcc actctacctc tgtccttcaa ggaagccact ctggccaaga ggtttgctct
     1981 gaagaccaga agtcagatca ctaagcggaa aaggatgtcc ctggtcaagg agaagaaagc
     2041 ggcccagacc ctcagtgcga tcttgcttgc cttcatcatc acttggaccc catacaacat
     2101 catggttctg gtgaacacct tttgtgacag ctgcataccc aaaacctttt ggaatctggg
     2161 ctactggctg tgctacatca acagcaccgt gaaccccgtg tgctatgctc tgtgcaacaa
     2221 aacattcaga accactttca agatgctgct gctgtgccag tgtgacaaaa aaaagaggcg
     2281 caagcagcag taccagcaga gacagtcggt catttttcac aagcgcgcac ccgagcaggc
     2341 cttgaaggat cccccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat
     2401 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2461 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2521 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2581 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     2641 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     2701 gacccaggtg agtacaggag gggtaccata acttcgtata aagtatccta tacgaagtta
     2761 tttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg
     2821 tataaggtat actatacgaa gttatgctag caaccccgac acttacctgt tggagcgttt
     2881 catgctttgc tgttgaagtt cgtaactgct gcagcttcga gaactgcccg tggggtggac
     2941 aaagttttct gtctctgcct ctgtcccaga ggcttgcagg ccagcaagct ctttggtacg
     3001 cttttcagtt tccttataga tcctccagta taaaatagtc ataatggtga caggcatata
     3061 aaaaccagcg atggctgtgc caaaagtaat ggtgggctca ctgaggaact gaatgaagca
     3121 ctctcccgga ggcacagttc tctttccaac aaagtattgc cagaacaaga tggcaggagc
     3181 ccaaaggaca aaggagatga cccaagccag accgatcatc acaccggctc tctttgttgt
     3241 tcgtttggct cggtacgtga gcggcctcgt gatggaaaag tatctgtcaa agctgatgac
     3301 cagaagattc ataacagagg cattgctggc tacgcagtca atggcaagcc agaggtcaca
     3361 ggccaagttc cctaaggccc atcgattcat gatgatgtag gtcgtaaaca gattcattga
     3421 aatgaccccg ataatcagat cggcacaggc caggcttaag aggaagtagt tgttgaccgt
     3481 cttcagctgc ttgttgacct taaatgacac aattaccagg atgttgccga tgatggtcac
     3541 caaggccagg atgcccgtta agaaagcgat gaagaccact tgccagacgg tatgacctcc
     3601 cagagggtca tcggtggtac cgtctggaga ggagaaattg ccagctgctc gagaaacatt
     3661 gtagctgccg aaatgagtga cggttcccgg gggcagccct gcatcggagg ggctgtgtat
     3721 ccaggaggag ctgatgtttg gaaacaaagg cgaggttgta ctgttattgt gcaaggtcat
     3781 ggtggcgagc tcgaagttcc tatactattt gaagaatagg aacttcttgc cttaacccag
     3841 aaattatcac tgttattctt tagaatggtg caaagagaag ttcctatacc ttttgaagaa
     3901 taggaacttc gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3961 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4021 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4081 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4141 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4201 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4261 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4321 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     4381 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4441 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4501 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     4561 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     4621 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     4681 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     4741 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     4801 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     4861 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4921 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4981 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     5041 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     5101 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5161 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5221 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5281 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     5341 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     5581 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     5641 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     5701 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     5761 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5821 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5881 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5941 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6001 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6061 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6121 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6181 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6241 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6301 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6361 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6421 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6481 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6541 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6601 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6661 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6721 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6781 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6841 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6901 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6961 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7021 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7801 tcacatgt
//
