LOCUS       Exported                6964 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV DREADD expression.
ACCESSION   .
VERSION     .
KEYWORDS    pZac2.1 GfaABC1D hM3D mCherry  SV40
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6964
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    101..122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        137..167
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    175..191
                     /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     180..202
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     199..215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     199..215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   253..382
                     /label=AAV2 ITR
     intron          1233..1365
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1348..1367
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1410..1428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3227..3236
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3233..3943
                     /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(3378..3396)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3646..3665
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3983..4104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3994..4013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4048..4067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4266..4395
                     /label=AAV2 ITR
     primer_bind     complement(4414..4431)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4414..4430)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4423..4445)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4572..5027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4659..4678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4869..4890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5053..5157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5158..6018
                     /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(5376..5395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6189..6777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6678..6697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6931..6948
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
       61 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      121 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      181 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga tttaattaag
      241 gccttaatta ggctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
      301 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      361 aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg
      421 tagccatgct ctaggaagat ctaacatatc ctggtgtgga gtaggggacg ctgctctgac
      481 agaggctcgg gggcctgagc tggctctgtg agctggggag gaggcagaca gccaggcctt
      541 gtctgcaagc agacctggca gcattgggct ggccgccccc cagggcctcc tcttcatgcc
      601 cagtgaatga ctcaccttgg cacagacaca atgttcgggg tgggcacagt gcctgcttcc
      661 cgccgcaccc cagcccccct caaatgcctt ccgagaagcc cattgagcag ggggcttgca
      721 ttgcacccca gcctgacagc ctggcatctt gggataaaag cagcacagcc ccctaggggc
      781 tgcccttgct gtgtggcgcc accggcggtg gagaacaagg ctctattcag cctgtgccca
      841 ggaaagggga tcaggggatg cccaggcatg gacagtgggt ggcagggggg gagaggaggg
      901 ctgtctgctt cccagaagtc caaggacaca aatgggtgag gggagagctc tccccatagc
      961 tgggctgcgg cccaacccca ccccctcagg ctatgccagg gggtgttgcc aggggcaccc
     1021 gggcatcgcc agtctagccc actccttcat aaagccctcg catcccagga gcgagcagag
     1081 ccagagcagg ttggagagga gacgcatcac ctccgctgct cgcaagcttt attgcggtag
     1141 tttatcacag ttaaattgct aacgcagtca gtgcttctga cacaacagtc tcgaacttaa
     1201 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     1261 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     1321 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagt
     1381 tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct agcctcgaga
     1441 attcaccatg accttgcaca ataacagtac aacctcgcct ttgtttccaa acatcagctc
     1501 ctcctggata cacagcccct ccgatgcagg gctgcccccg ggaaccgtca ctcatttcgg
     1561 cagctacaat gtttctcgag cagctggcaa tttctcctct ccagacggta ccaccgatga
     1621 ccctctggga ggtcataccg tctggcaagt ggtcttcatc gctttcttaa cgggcatcct
     1681 ggccttggtg accatcatcg gcaacatcct ggtaattgtg tcatttaagg tcaacaagca
     1741 gctgaagacg gtcaacaact acttcctctt aagcctggcc tgtgccgatc tgattatcgg
     1801 ggtcatttca atgaatctgt ttacgaccta catcatcatg aatcgatggg ccttagggaa
     1861 cttggcctgt gacctctggc ttgccattga ctgcgtagcc agcaatgcct ctgttatgaa
     1921 tcttctggtc atcagctttg acagatactt ttccatcacg aggccgctca cgtaccgagc
     1981 caaacgaaca acaaagagag ccggtgtgat gatcggtctg gcttgggtca tctcctttgt
     2041 cctttgggct cctgccatct tgttctggca atactttgtt ggaaagagaa ctgtgcctcc
     2101 gggagagtgc ttcattcagt tcctcagtga gcccaccatt acttttggca cagccatcgc
     2161 tggtttttat atgcctgtca ccattatgac tattttatac tggaggatct ataaggaaac
     2221 tgaaaagcgt accaaagagc ttgctggcct gcaagcctct gggacagagg cagagacaga
     2281 aaactttgtc caccccacgg gcagttctcg aagctgcagc agttacgaac ttcaacagca
     2341 aagcatgaaa cgctccaaca ggaggaagta tggccgctgc cacttctggt tcacaaccaa
     2401 gagctggaaa cccagctccg agcagatgga ccaagaccac agcagcagtg acagttggaa
     2461 caacaatgat gctgctgcct ccctggagaa ctccgcctcc tccgacgagg aggacattgg
     2521 ctccgagacg agagccatct actccatcgt gctcaagctt ccgggtcaca gcaccatcct
     2581 caactccacc aagttaccct catcggacaa cctgcaggtg cctgaggagg agctggggat
     2641 ggtggacttg gagaggaaag ccgacaagct gcaggcccag aagagcgtgg acgatggagg
     2701 cagttttcca aaaagcttct ccaagcttcc catccagcta gagtcagccg tggacacagc
     2761 taagacttct gacgtcaact cctcagtggg taagagcacg gccactctac ctctgtcctt
     2821 caaggaagcc actctggcca agaggtttgc tctgaagacc agaagtcaga tcactaagcg
     2881 gaaaaggatg tccctggtca aggagaagaa agcggcccag accctcagtg cgatcttgct
     2941 tgccttcatc atcacttgga ccccatacaa catcatggtt ctggtgaaca ccttttgtga
     3001 cagctgcata cccaaaacct tttggaatct gggctactgg ctgtgctaca tcaacagcac
     3061 cgtgaacccc gtgtgctatg ctctgtgcaa caaaacattc agaaccactt tcaagatgct
     3121 gctgctgtgc cagtgtgaca aaaaaaagag gcgcaagcag cagtaccagc agagacagtc
     3181 ggtcattttt cacaagcgcg cacccgagca ggccttggga ccggtcgcca ccatggtgag
     3241 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     3301 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3361 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     3421 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     3481 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3541 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     3601 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     3661 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     3721 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     3781 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     3841 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     3901 gggccgccac tccaccggcg gcatggacga gctgtacaag tagtctagag tcgacccggg
     3961 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4021 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4081 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4141 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     4201 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     4261 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4321 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4381 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
     4441 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4501 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4561 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4921 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4981 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5101 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5161 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5221 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5281 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5341 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5401 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5461 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5521 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5581 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5641 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5701 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5761 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5821 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5881 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5941 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6001 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6061 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6121 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6301 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6541 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6961 cgcc
//
