LOCUS       sequence-177827-        6619 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 6619)
  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 6619)
  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 hM4D mCherry SV40
FEATURES             Location/Qualifiers
     source          1..6619
                     /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      2891..2900
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2897..3604
                     /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(3042..3060)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3310..3329
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3644..3765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3655..3674)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3709..3728
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3927..4056
                     /label=AAV2 ITR
     primer_bind     complement(4075..4092)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4075..4091)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4084..4106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4233..4688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4320..4339)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4530..4551
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4819..4887,4888..5679)
                     /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      5850..6438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6339..6358
                     /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 taggctagat
     1441 ggccaacttc acacctgtca atggcagctc gggcaatcag tccgtgcgcc tggtcacgtc
     1501 atcatcccac aatcgctatg agacggtgga aatggtcttc attgccacag tgacaggctc
     1561 cctgagcctg gtgactgtcg tgggcaacat cctggtgatg ctgtccatca aggtcaacag
     1621 gcagctgcag acagtcaaca actacttcct cttcagcctg gcgtgtgctg atctcatcat
     1681 aggcgccttc tccatgaacc tctacaccgt gtacatcatc aagggctact ggcccctggg
     1741 cgccgtggtc tgcgacctgt ggctggccct ggactgcgtg gtgagcaacg cctccgtcat
     1801 gaaccttctc atcatcagct ttgaccgcta cttctgcgtc accaagcctc tcacctaccc
     1861 tgcccggcgc accaccaaga tggcaggcct catgattgct gctgcctggg tactgtcctt
     1921 cgtgctctgg gcgcctgcca tcttgttctg gcagtttgtg gtgggtaagc ggacggtgcc
     1981 cgacaaccag tgcttcatcc agttcctgtc caacccagca gtgacctttg gcacagccat
     2041 tgctggcttc tacctgcctg tggtcatcat gacggtgctg tacatccaca tctccctggc
     2101 cagtcgcagc cgagtccaca agcaccggcc cgagggcccg aaggagaaga aagccaagac
     2161 gctggccttc ctcaagagcc cactaatgaa gcagagcgtc aagaagcccc cgcccgggga
     2221 ggccgcccgg gaggagctgc gcaatggcaa gctggaggag gcccccccgc cagcgctgcc
     2281 accgccaccg cgccccgtgg ctgataagga cacttccaat gagtccagct caggcagtgc
     2341 cacccagaac accaaggaac gcccagccac agagctgtcc accacagagg ccaccacgcc
     2401 cgccatgccc gcccctcccc tgcagccgcg ggccctcaac ccagcctcca gatggtccaa
     2461 gatccagatt gtgacgaagc agacaggcaa tgagtgtgtg acagccattg agattgtgcc
     2521 tgccacgccg gctggcatgc gccctgcggc caacgtggcc cgcaagttcg ccagcatcgc
     2581 tcgcaaccag gtgcgcaaga agcggcagat ggcggcccgg gagcgcaaag tgacacgaac
     2641 gatctttgcc attctgctgg ccttcatcct cacctggacg ccctacaacg tcatggtcct
     2701 ggtgaacacc ttctgccaga gctgcatccc tgacacggtg tggtccattg gctactggct
     2761 ctgctacgtc aacagcacca tcaaccctgc ctgctatgct ctgtgcaacg ccacctttaa
     2821 aaagaccttc cggcacctgc tgctgtgcca gtatcggaac atcggcactg ccaggcggga
     2881 tccaccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     2941 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3001 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3061 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     3121 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     3181 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3241 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     3301 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     3361 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     3421 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     3481 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     3541 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3601 caagtctaga gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     3661 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3721 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3781 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3841 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     3901 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     3961 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4021 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4081 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4141 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4201 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4261 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4321 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4381 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4441 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4501 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4561 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4621 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4681 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4741 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4801 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4861 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4921 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4981 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5041 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5101 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5161 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5221 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5281 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5341 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5401 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5461 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5521 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5581 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5641 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5701 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5761 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5821 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5881 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5941 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6001 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6061 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6121 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6181 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6241 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6301 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6361 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6421 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6481 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6541 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6601 gtgagcgagg aagcggaag
//
