LOCUS       sequence-212680-        8936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a plasma membrane calcium pump hPMCA2x/b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8936)
  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 8936)
  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-hPMCA2x/b
FEATURES             Location/Qualifiers
     source          1..8936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          846..978
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     961..980
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1023..1041
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1047..1754
                     /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(1192..1210)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1460..1479
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1770..1796
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1797..1814
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1815..1832
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      1884..1893
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2391..2402)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2859..2868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    5568..5689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5579..5598)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5633..5652
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5851..5980
                     /label=AAV2 ITR
     primer_bind     complement(5999..6016)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5999..6015)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6008..6030)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6157..6612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6244..6263)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6454..6475
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6638..6742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6743..6811,6812..7603)
                     /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      7774..8362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8263..8282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    8650..8671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8686..8691,8692..8709,8710..8716)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8724..8740
                     /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     8729..8751
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8748..8764
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8748..8764
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   8802..8931
                     /label=AAV2 ITR
ORIGIN
        1 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatctaacat
       61 atcctggtgt ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct
      121 gtgagctggg gaggaggcag acagccaggc cttgtctgca agcagacctg gcagcattgg
      181 gctggccgcc ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac
      241 acaatgttcg gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc
      301 cttccgagaa gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat
      361 cttgggataa aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg
      421 gtggagaaca aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc
      481 atggacagtg ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac
      541 acaaatgggt gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc
      601 aggctatgcc agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt
      661 cataaagccc tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat
      721 cacctccgct gctcgcaagc tttattgcgg tagtttatca cagttaaatt gctaacgcag
      781 tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg tgaggcactg
      841 ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt
      901 cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact
      961 ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta
     1021 cttaatacga ctcactatag gctagcatgg tgagcaaggg cgaggaggat aacatggcca
     1081 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1141 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1201 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     1261 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     1321 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     1381 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1441 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1501 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1561 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1621 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1681 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1741 acgagctgta caagtccgga ctcagatcct acccctatga tgtgcctgac tatgcccacc
     1801 accaccacca ccacctggtg cccaggggca gcagatctcg agctcaagct tcgaattctg
     1861 cagtcgacgg taccgcggtg gcggccgcca tgggtgacat gaccaacagc gacttttact
     1921 ccaaaaacca aagaaatgag tcgagccatg ggggcgagtt cgggtgcaca atggaggagc
     1981 tccgctccct catggagctg cggggcactg aggctgtggt caagatcaag gagacttatg
     2041 gggacaccga agccatctgc cggcgcctca aaacctcacc tgttgaaggt ttgccggaca
     2101 ccgctccaga cctggaaaag agaaagcaaa tttttgggca aaactttata cctccaaaga
     2161 agccaaaaac cttcctgcag ctcgtgtggg aggcgctgca ggacgtgacg ctcatcatcc
     2221 tggagattgc cgccatcatc tccctggggc tgtccttcta ccacccgccc ggcgagggca
     2281 acgaaggatg tgcgacggcc cagggtgggg cagaggatga aggagaggca gaggcaggtt
     2341 ggatcgaggg ggccgccatt ctcctctcag ttatctgtgt ggtcctggtc acggccttca
     2401 atgactggag caaagagaaa cagttccggg gcctgcagag ccgcatcgag caggaacaga
     2461 aatttaccgt agtccgggct ggccaggtgg tgcagatccc tgtggctgag atcgtggttg
     2521 gggacatagc ccaggtcaaa tatggtgacc tcctccctgc cgacggcctc ttcatccagg
     2581 gcaatgacct caagattgat gaaagctccc taactggaga gtctgaccag gtgcgcaagt
     2641 ccgtggacaa ggaccccatg ctgctgtcag gaacccacgt gatggagggc tcaggacgga
     2701 tgttggtgac tgctgtgggt gtgaactctc agactggcat catctttacc ctcctggggg
     2761 ctggtggtga agaggaagag aagaaagaca aaaaaggtaa aatgcaggat ggcaatgtgg
     2821 acgccagcca gagcaaagcc aaacaacagg acggggcagc cgccatggag atgcagcccc
     2881 tcaagagtgc cgagggcggc gacgctgacg acaggaagaa ggccagcatg cacaagaagg
     2941 agaagtccgt gctgcagggc aagctcacca agctggctgt gcagatcggg aaggcgggct
     3001 tggtgatgtc agccatcacg gtgatcatcc tggtgctcta cttcactgtg gacaccttcg
     3061 tggtcaacaa gaagccgtgg ctgcctgagt gcacgcccgt ctacgtgcag tactttgtca
     3121 agttcttcat cattggcgtg acggtgctgg tggtcgccgt gcccgagggg ctccctctgg
     3181 ccgtcaccat ctcgttggcc tattcggtga agaaaatgat gaaggacaac aacctggtac
     3241 gccacctgga tgcctgtgag accatgggca atgccacagc catctgctca gacaagacag
     3301 gcacgctgac caccaatcgc atgacagtgg tacaggccta tgtcggcgac gtccactata
     3361 aagagatccc cgaccccagc tccatcaaca ccaagaccat ggagctgctg atcaatgcca
     3421 tcgccatcaa cagcgcctac accaccaaga ttctgccccc agagaaggag ggcgccctgc
     3481 ctcggcaggt gggcaacaag acggagtgcg gcctgctggg cttcgtgctg gacctgaagc
     3541 aggactacga gcccgtgcgc agccagatgc cagaggagaa gttgtacaaa gtgtacacct
     3601 tcaactccgt gcgcaagtcc atgagcactg tcatcaagct gcccgacgag agcttccgca
     3661 tgtacagcaa gggggcttct gagatcgtgc tcaagaagtg ctgcaaaatc ctcaatgggg
     3721 cgggagagcc tcgtgtcttc cggccccgcg accgggacga gatggtaaag aaggtgattg
     3781 agcccatggc ttgcgatggg ctccgcacta tctgcgtggc ctaccgcgac ttccccagca
     3841 gcccggagcc ggactgggac aatgagaatg acatcctcaa cgaactcacc tgcatctgcg
     3901 tggtgggcat cgaggacccg gtgcggccag aggtcccaga agccatccgc aagtgccagc
     3961 gggcaggcat cacggtccgc atggtcactg gcgacaatat caacacggct cgggccatcg
     4021 ccatcaagtg tggcatcatc catcctgggg aggactttct gtgcctcgag ggcaaggagt
     4081 tcaacaggag gatccgcaac gagaaggggg agattgagca ggagcgaatt gacaagatct
     4141 ggccaaagct gcgggtgctg gctcgctcct ccccaacgga caagcatacc ctggttaaag
     4201 gcatcatcga cagcacacac actgagcagc ggcaggtggt ggccgtgacg ggggacggga
     4261 ccaacgacgg gcctgcactc aagaaggccg acgtgggctt cgccatgggc atcgcaggca
     4321 ctgacgtggc caaggaggcc tcagacatca tcctgacaga cgacaatttc agcagcatcg
     4381 tcaaggcagt gatgtggggc cgcaacgtct atgacagcat ctccaaattc ttgcagttcc
     4441 agctcaccgt caacgtggtg gccgtgattg tggccttcac aggcgcctgc atcacgcagg
     4501 actcccctct gaaggccgtg cagatgctct gggtgaacct catcatggac acgtttgcct
     4561 cgctggcact ggccactgag ccgcccacgg agaccctgct gctgaggaag ccgtacggcc
     4621 gcaacaagcc gctcatctcc aggaccatga tgaagaacat cctgggccat gctgtctacc
     4681 agcttgccct catcttcacc ctgctctttg ttggcgagaa gatgttccag atcgacagcg
     4741 ggaggaacgc gcccctgcat tcgccaccct cagaacatta caccatcatc ttcaacacct
     4801 tcgtcatgat gcagctcttc aacgagatca acgcccgcaa gatccacggc gagcgcaatg
     4861 tctttgacgg catcttccgg aaccccatct tctgcaccat cgtgctgggc acctttgcca
     4921 tccagatagt gatcgtgcag tttggaggga agccattcag ctgctctcca ctgcagctgg
     4981 accagtggat gtggtgcata ttcattgggt taggagagct cgtttggggc caggtcatcg
     5041 ccaccatccc gaccagcaga ctcaagttcc tcaaggaggc aggcaggctc acacagaagg
     5101 aggagatccc ggaggaggag ctcaacgagg acgtggagga gatcgaccac gcggagcggg
     5161 agctgcggcg gggccagatc ctgtggttcc gaggcctgaa tcggatccag acacagatcc
     5221 gcgtcgtgaa ggcgttccgt agctctctct atgaaggttt agaaaagcct gaatctcgaa
     5281 cctccatcca taacttcatg gctcatcctg aattccggat cgaagattcc cagccccaca
     5341 tccccctcat tgatgacacc gacctggaag aagatgccgc gctcaagcag aactcgagcc
     5401 cgccgtcatc cctcaacaag aacaacagcg ccatcgacag tgggatcaac ctgacgaccg
     5461 acacaagcaa atcagctacc tcttcaagtc cagggagccc catccacagc ctggagacgt
     5521 cgctttagtc tagagtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga
     5581 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5641 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5701 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     5761 aaacctctac aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata
     5821 agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc
     5881 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5941 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa cctaattcac
     6001 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6061 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6121 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     6181 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6241 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6301 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6361 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6421 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6481 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6541 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6601 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     6901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     6961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     7741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     7801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     7861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     7921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     7981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     8101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     8161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     8221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     8281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     8341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     8401 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     8461 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     8521 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     8581 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     8641 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     8701 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     8761 tgaccatgat tacgccagat ttaattaagg ccttaattag gctgcgcgct cgctcgctca
     8821 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     8881 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtag
//
