LOCUS       sequence-212681-        9029 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a plasma membrane calcium pump hPMCA2w/b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9029)
  AUTHORS   Yu X, Taylor AMW, Nagai J, Golshani P, Evans CJ, Coppola G, Khakh BS
  TITLE     Reducing Astrocyte Calcium Signaling In Vivo Alters Striatal 
            Microcircuits and Causes Repetitive Behavior.
  JOURNAL   Neuron. 2018 Aug 27. pii: S0896-6273(18)30688-3. doi: 
            10.1016/j.neuron.2018.08.015.
  PUBMED    30174118
REFERENCE   2  (bases 1 to 9029)
  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"; journalName: "Neuron. 
            2018 Aug 27. pii: S0896-6273(18)30688-3. doi: 
            10.1016/j.neuron.2018.08.015."
COMMENT     Sequence Label: pZac2.1-GfaABC1D-mCherry-hPMCA2w/b
FEATURES             Location/Qualifiers
     source          1..9029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   41..170
                     /label=AAV2 ITR
     intron          1021..1153
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1136..1155
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1198..1216
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1222..1929
                     /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(1367..1385)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1635..1654
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1945..1971
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1972..1989
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1990..2007
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      2059..2068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2566..2577)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3127..3136
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5836..5957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5847..5866)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5901..5920
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6119..6248
                     /label=AAV2 ITR
     primer_bind     complement(6267..6284)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6267..6283)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6276..6298)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6425..6880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6512..6531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6722..6743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6906..7010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7011..7079,7080..7871)
                     /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      8042..8630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8531..8550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    8918..8939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8954..8959,8960..8977,8978..8984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8992..9008
                     /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     8997..9019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(9016..9029,1..3)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(9016..9029,1..3)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaccatgatt acgccagatt taattaaggc cttaattagg ctgcgcgctc gctcgctcac
       61 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
      121 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat
      181 gattaacccg ccatgctact tatctacgta gccatgctct aggaagatct aacatatcct
      241 ggtgtggagt aggggacgct gctctgacag aggctcgggg gcctgagctg gctctgtgag
      301 ctggggagga ggcagacagc caggccttgt ctgcaagcag acctggcagc attgggctgg
      361 ccgcccccca gggcctcctc ttcatgccca gtgaatgact caccttggca cagacacaat
      421 gttcggggtg ggcacagtgc ctgcttcccg ccgcacccca gcccccctca aatgccttcc
      481 gagaagccca ttgagcaggg ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg
      541 gataaaagca gcacagcccc ctaggggctg cccttgctgt gtggcgccac cggcggtgga
      601 gaacaaggct ctattcagcc tgtgcccagg aaaggggatc aggggatgcc caggcatgga
      661 cagtgggtgg caggggggga gaggagggct gtctgcttcc cagaagtcca aggacacaaa
      721 tgggtgaggg gagagctctc cccatagctg ggctgcggcc caaccccacc ccctcaggct
      781 atgccagggg gtgttgccag gggcacccgg gcatcgccag tctagcccac tccttcataa
      841 agccctcgca tcccaggagc gagcagagcc agagcaggtt ggagaggaga cgcatcacct
      901 ccgctgctcg caagctttat tgcggtagtt tatcacagtt aaattgctaa cgcagtcagt
      961 gcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag
     1021 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
     1081 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     1141 tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta gagtacttaa
     1201 tacgactcac tataggctag catggtgagc aagggcgagg aggataacat ggccatcatc
     1261 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     1321 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     1381 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     1441 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1501 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1561 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1621 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1681 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1741 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1801 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     1861 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     1921 ctgtacaagt ccggactcag atcctacccc tatgatgtgc ctgactatgc ccaccaccac
     1981 caccaccacc tggtgcccag gggcagcaga tctcgagctc aagcttcgaa ttctgcagtc
     2041 gacggtaccg cggtggcggc cgccatgggt gacatgacca acagcgactt ttactccaaa
     2101 aaccaaagaa atgagtcgag ccatgggggc gagttcgggt gcacaatgga ggagctccgc
     2161 tccctcatgg agctgcgggg cactgaggct gtggtcaaga tcaaggagac ttatggggac
     2221 accgaagcca tctgccggcg cctcaaaacc tcacctgttg aaggtttgcc ggacaccgct
     2281 ccagacctgg aaaagagaaa gcaaattttt gggcaaaact ttatacctcc aaagaagcca
     2341 aaaaccttcc tgcagctcgt gtgggaggcg ctgcaggacg tgacgctcat catcctggag
     2401 attgccgcca tcatctccct ggggctgtcc ttctaccacc cgcccggcga gggcaacgaa
     2461 ggatgtgcga cggcccaggg tggggcagag gatgaaggag aggcagaggc aggttggatc
     2521 gagggggccg ccattctcct ctcagttatc tgtgtggtcc tggtcacggc cttcaatgac
     2581 tggagcaaag agaaacagtt ccggggcctg cagagccgca tcgagcagga acagaaattt
     2641 accgtagtcc gggctggcca ggtggtgcag atccctgtgg ctgagatcgt ggttggggac
     2701 atagcccagg tcaaatatgg tgacctcctc cctgccgacg gcctcttcat ccagggcaat
     2761 gacctcaaga ttgatgaaag ctccctaact ggagagtctg accaggtgcg caagtccgtg
     2821 gacaaggacc ccatgctgct gtcaggaacc cacgtgatgg agggctcagg acggatgttg
     2881 gtgactgctg tgggtgtgaa ctctcagact ggcatcatct ttaccctcct gggggctggt
     2941 ggtgaagagg aagagaagaa agacaaaaaa ggtgtgaaga agggggatgg ccttcagcta
     3001 ccagcagcag acggtgcggc agcttcaaat gctgcagata gtgcgaatgc cagcctagtc
     3061 aatggtaaaa tgcaggatgg caatgtggac gccagccaga gcaaagccaa acagcaggac
     3121 ggggcagccg ccatggagat gcagcccctc aagagtgccg agggcggcga cgctgacgac
     3181 aggaagaagg ccagcatgca caagaaggag aagtccgtgc tgcagggcaa gctcaccaag
     3241 ctggctgtgc agatcgggaa ggcgggcttg gtgatgtcag ccatcacggt gatcatcctg
     3301 gtgctctact tcactgtgga caccttcgtg gtcaacaaga agccgtggct gcctgagtgc
     3361 acgcccgtct acgtgcagta ctttgtcaag ttcttcatca ttggcgtgac ggtgctggtg
     3421 gtcgccgtgc ccgaggggct ccctctggcc gtcaccatct cgttggccta ttcggtgaag
     3481 aaaatgatga aggacaacaa cctggtacgc cacctggatg cctgtgagac catgggcaat
     3541 gccacagcca tctgctcaga caagacaggc acgctgacca ccaatcgcat gacagtggta
     3601 caggcctatg tcggcgacgt ccactataaa gagatccccg accccagctc catcaacacc
     3661 aagaccatgg agctgctgat caatgccatc gccatcaaca gcgcctacac caccaagatt
     3721 ctgcccccag agaaggaggg cgccctgcct cggcaggtgg gcaacaagac ggagtgcggc
     3781 ctgctgggct tcgtgctgga cctgaagcag gactacgagc ccgtgcgcag ccagatgcca
     3841 gaggagaagt tgtacaaagt gtacaccttc aactccgtgc gcaagtccat gagcactgtc
     3901 atcaagctgc ccgacgagag cttccgcatg tacagcaagg gggcttctga gatcgtgctc
     3961 aagaagtgct gcaaaatcct caatggggcg ggagagcctc gtgtcttccg gccccgcgac
     4021 cgggacgaga tggtaaagaa ggtgattgag cccatggctt gcgatgggct ccgcactatc
     4081 tgcgtggcct accgcgactt ccccagcagc ccggagccgg actgggacaa tgagaatgac
     4141 atcctcaacg aactcacctg catctgcgtg gtgggcatcg aggacccggt gcggccagag
     4201 gtcccagaag ccatccgcaa gtgccagcgg gcaggcatca cggtccgcat ggtcactggc
     4261 gacaatatca acacggctcg ggccatcgcc atcaagtgtg gcatcatcca tcctggggag
     4321 gactttctgt gcctcgaggg caaggagttc aacaggagga tccgcaacga gaagggggag
     4381 attgagcagg agcgaattga caagatctgg ccaaagctgc gggtgctggc tcgctcctcc
     4441 ccaacggaca agcataccct ggttaaaggc atcatcgaca gcacacacac tgagcagcgg
     4501 caggtggtgg ccgtgacggg ggacgggacc aacgacgggc ctgcactcaa gaaggccgac
     4561 gtgggcttcg ccatgggcat cgcaggcact gacgtggcca aggaggcctc agacatcatc
     4621 ctgacagacg acaatttcag cagcatcgtc aaggcagtga tgtggggccg caacgtctat
     4681 gacagcatct ccaaattctt gcagttccag ctcaccgtca acgtggtggc cgtgattgtg
     4741 gccttcacag gcgcctgcat cacgcaggac tcccctctga aggccgtgca gatgctctgg
     4801 gtgaacctca tcatggacac gtttgcctcg ctggcactgg ccactgagcc gcccacggag
     4861 accctgctgc tgaggaagcc gtacggccgc aacaagccgc tcatctccag gaccatgatg
     4921 aagaacatcc tgggccatgc tgtctaccag cttgccctca tcttcaccct gctctttgtt
     4981 ggcgagaaga tgttccagat cgacagcggg aggaacgcgc ccctgcattc gccaccctca
     5041 gaacattaca ccatcatctt caacaccttc gtcatgatgc agctcttcaa cgagatcaac
     5101 gcccgcaaga tccacggcga gcgcaatgtc tttgacggca tcttccggaa ccccatcttc
     5161 tgcaccatcg tgctgggcac ctttgccatc cagatagtga tcgtgcagtt tggagggaag
     5221 ccattcagct gctctccact gcagctggac cagtggatgt ggtgcatatt cattgggtta
     5281 ggagagctcg tttggggcca ggtcatcgcc accatcccga ccagcagact caagttcctc
     5341 aaggaggcag gcaggctcac acagaaggag gagatcccgg aggaggagct caacgaggac
     5401 gtggaggaga tcgaccacgc ggagcgggag ctgcggcggg gccagatcct gtggttccga
     5461 ggcctgaatc ggatccagac acagatccgc gtcgtgaagg cgttccgtag ctctctctat
     5521 gaaggtttag aaaagcctga atctcgaacc tccatccata acttcatggc tcatcctgaa
     5581 ttccggatcg aagattccca gccccacatc cccctcattg atgacaccga cctggaagaa
     5641 gatgccgcgc tcaagcagaa ctcgagcccg ccgtcatccc tcaacaagaa caacagcgcc
     5701 atcgacagtg ggatcaacct gacgaccgac acaagcaaat cagctacctc ttcaagtcca
     5761 gggagcccca tccacagcct ggagacgtcg ctttagtcta gagtcgaccc gggcggccgc
     5821 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5881 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5941 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     6001 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     6061 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     6121 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     6181 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     6241 gcgcgcagcc ttaattaacc taattcactg gccgtcgttt tacaacgtcg tgactgggaa
     6301 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     6361 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     6421 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6481 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6541 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6601 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6661 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6721 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6781 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6841 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     6901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7081 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7141 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7201 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7261 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     7321 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     7381 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7441 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7501 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7561 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7621 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7681 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     7741 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7801 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7861 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8161 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     8281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     8341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     8401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     8761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     8821 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     8881 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     8941 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     9001 gataacaatt tcacacagga aacagctat
//
