LOCUS       sequence-177824-        6434 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 6434)
  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 6434)
  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 rM3D mCherry SV40
FEATURES             Location/Qualifiers
     source          1..6434
                     /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      2703..2712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2709..3419
                     /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(2854..2872)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3122..3141
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3459..3580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3470..3489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3524..3543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3742..3871
                     /label=AAV2 ITR
     primer_bind     complement(3890..3907)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3890..3906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3899..3921)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4048..4503
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4135..4154)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4345..4366
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4529..4633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4634..4702,4703..5494)
                     /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      5665..6253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6154..6173
                     /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 tcgaggccac catgaccttg cacagtaaca gtacaacctc gcctttgttt cccaacatca
     1501 gctcttcctg ggtgcacagt ccctcggagg cagggctgcc cttggggaca gtcactcagt
     1561 tgggcagcta caacatttca caagaaactg ggaatttctc ctcaaacgac acctccagcg
     1621 accctctcgg gggtcacacc atctggcaag tggtcttcat tgccttctta actggcttcc
     1681 tggcattggt gaccatcatt ggcaacatcc ttgtcattgt ggccttcaag gtcaacaaac
     1741 agctgaagac agtcaacaac tacttcctct taagcctggc ctgtgcagac ctgatcatcg
     1801 gggtcatttc catgaacctg ttcactacct acatcattat gaaccgttgg gcactgggga
     1861 acttagcctg cgacctctgg ctctccattg actgtgtggc cagcaatgcc tctgtcatga
     1921 atctgctggt catcagcttt gacaggtact tttccatcac ttctccattc cgctaccaga
     1981 gcctgatgac cagggctcga gctggtgtga tgattggtct ggcttgggtc atctcctttg
     2041 tcctatgggc tcctgccatc ttgttctggc aatactttgt agggaagaga actgtgcccc
     2101 caggagaatg tttcattcag tttctgagtg agcccaccat caccttcggc acggcgatcg
     2161 ctggctttta catgcctgtc accatcatga ctattttata ctggcgggtg taccgggagg
     2221 ccaaggagca gatcaggaag atcgaccgct gcgagggccg gttctatggc agccaggagc
     2281 agccgcagcc acccccgctc ccccaacacc agcccatcct cggcaacggc cgtgccagca
     2341 agaggaagac gtcccgtgtc atggccatga gggaacacaa agctctgcag acgctcagtg
     2401 ccatcttgct ggccttcatc atcacgtgga ccccctacaa catcatggtc ctggtgaaca
     2461 ccttctgtga cagctgcata cccaaaacct attggaatct gggctactgg ctgtgctata
     2521 tcaacagcac cgtgaaccct gtgtgctatg ccctgtgcaa caaaacattc agaaccacct
     2581 tcaagacgct cctcttgtgc cagtgtgaca aaaggaagag gcgcaaacag cagtaccagc
     2641 agagacagtc ggtcattttt cacaagcgag tgccggagca ggccttggcg gatccaccgg
     2701 tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     2761 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2821 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2881 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     2941 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     3001 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     3061 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     3121 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     3181 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     3241 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     3301 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     3361 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaat
     3421 ctagagtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3481 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3541 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3601 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     3661 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     3721 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3781 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3841 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     3901 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3961 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4021 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4081 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4141 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4201 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4261 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4321 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4381 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4441 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4501 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4561 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4621 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4681 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4741 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4801 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4861 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4921 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4981 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5041 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5101 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5161 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5221 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5281 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5341 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     5401 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     5461 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     5521 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6361 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6421 cgaggaagcg gaag
//
