LOCUS       sequence-177823-        6964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV DREADD expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6964)
  AUTHORS   Chai H, Diaz-Castro B, Shigetomi E, Monte E, Octeau JC, Yu X, Cohn 
            W, Rajendran PS, Vondriska TM, Whitelegge JP, Coppola G, Khakh BS
  TITLE     Neural Circuit-Specialized Astrocytes: Transcriptomic, Proteomic, 
            Morphological, and Functional Evidence.
  JOURNAL   Neuron. 2017 Aug 2;95(3):531-549.e9. doi: 
            10.1016/j.neuron.2017.06.029. Epub 2017 Jul 14.
  PUBMED    28712653
REFERENCE   2  (bases 1 to 6964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2017.06.029"; journalName: "Neuron"; date: 
            "2017-08-2- 2"; volume: "95"; issue: "3"; pages: "531-549.e9"
COMMENT     Sequence Label: pZac2.1 GfaABC1D hM3D mCherry  SV40
FEATURES             Location/Qualifiers
     source          1..6964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   259..388
                     /label=AAV2 ITR
     intron          1239..1371
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1354..1373
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1416..1434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3233..3242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3239..3949
                     /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(3384..3402)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3652..3671
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3989..4110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4000..4019)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4054..4073
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4272..4401
                     /label=AAV2 ITR
     primer_bind     complement(4420..4437)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4420..4436)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4429..4451)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4578..5033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4665..4684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4875..4896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5059..5163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5164..5232,5233..6024)
                     /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"
     rep_origin      6195..6783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6684..6703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccagattta
      241 attaaggcct taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
      301 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      361 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      421 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
      481 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
      541 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
      601 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
      661 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
      721 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
      781 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
      841 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
      901 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
      961 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1021 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1081 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1141 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     1201 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     1261 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     1321 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     1381 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     1441 tcgagaattc accatgacct tgcacaataa cagtacaacc tcgcctttgt ttccaaacat
     1501 cagctcctcc tggatacaca gcccctccga tgcagggctg cccccgggaa ccgtcactca
     1561 tttcggcagc tacaatgttt ctcgagcagc tggcaatttc tcctctccag acggtaccac
     1621 cgatgaccct ctgggaggtc ataccgtctg gcaagtggtc ttcatcgctt tcttaacggg
     1681 catcctggcc ttggtgacca tcatcggcaa catcctggta attgtgtcat ttaaggtcaa
     1741 caagcagctg aagacggtca acaactactt cctcttaagc ctggcctgtg ccgatctgat
     1801 tatcggggtc atttcaatga atctgtttac gacctacatc atcatgaatc gatgggcctt
     1861 agggaacttg gcctgtgacc tctggcttgc cattgactgc gtagccagca atgcctctgt
     1921 tatgaatctt ctggtcatca gctttgacag atacttttcc atcacgaggc cgctcacgta
     1981 ccgagccaaa cgaacaacaa agagagccgg tgtgatgatc ggtctggctt gggtcatctc
     2041 ctttgtcctt tgggctcctg ccatcttgtt ctggcaatac tttgttggaa agagaactgt
     2101 gcctccggga gagtgcttca ttcagttcct cagtgagccc accattactt ttggcacagc
     2161 catcgctggt ttttatatgc ctgtcaccat tatgactatt ttatactgga ggatctataa
     2221 ggaaactgaa aagcgtacca aagagcttgc tggcctgcaa gcctctggga cagaggcaga
     2281 gacagaaaac tttgtccacc ccacgggcag ttctcgaagc tgcagcagtt acgaacttca
     2341 acagcaaagc atgaaacgct ccaacaggag gaagtatggc cgctgccact tctggttcac
     2401 aaccaagagc tggaaaccca gctccgagca gatggaccaa gaccacagca gcagtgacag
     2461 ttggaacaac aatgatgctg ctgcctccct ggagaactcc gcctcctccg acgaggagga
     2521 cattggctcc gagacgagag ccatctactc catcgtgctc aagcttccgg gtcacagcac
     2581 catcctcaac tccaccaagt taccctcatc ggacaacctg caggtgcctg aggaggagct
     2641 ggggatggtg gacttggaga ggaaagccga caagctgcag gcccagaaga gcgtggacga
     2701 tggaggcagt tttccaaaaa gcttctccaa gcttcccatc cagctagagt cagccgtgga
     2761 cacagctaag acttctgacg tcaactcctc agtgggtaag agcacggcca ctctacctct
     2821 gtccttcaag gaagccactc tggccaagag gtttgctctg aagaccagaa gtcagatcac
     2881 taagcggaaa aggatgtccc tggtcaagga gaagaaagcg gcccagaccc tcagtgcgat
     2941 cttgcttgcc ttcatcatca cttggacccc atacaacatc atggttctgg tgaacacctt
     3001 ttgtgacagc tgcataccca aaaccttttg gaatctgggc tactggctgt gctacatcaa
     3061 cagcaccgtg aaccccgtgt gctatgctct gtgcaacaaa acattcagaa ccactttcaa
     3121 gatgctgctg ctgtgccagt gtgacaaaaa aaagaggcgc aagcagcagt accagcagag
     3181 acagtcggtc atttttcaca agcgcgcacc cgagcaggcc ttgggaccgg tcgccaccat
     3241 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3301 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3361 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3421 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3481 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3541 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3601 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3661 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3721 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3781 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3841 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3901 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtagt ctagagtcga
     3961 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     4021 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     4081 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     4141 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     4201 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     4261 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4321 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4381 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     4441 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4501 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     4561 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4621 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4681 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4741 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4801 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4861 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4921 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4981 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5041 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5101 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5161 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5221 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5281 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5341 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5401 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5461 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5521 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5581 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5641 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5701 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5761 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5821 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5881 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5941 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6001 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6061 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6121 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6181 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6241 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6301 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6361 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6421 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6481 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6541 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6601 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6661 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6721 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6781 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6841 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6901 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6961 gaag
//
