LOCUS       sequence-217178-        4955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Rpl22HA in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4955)
  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 4955)
  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-Rpl22-HA
FEATURES             Location/Qualifiers
     source          1..4955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    262..283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(298..303,304..321,322..328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    336..352
                     /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     341..363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     360..376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     360..376
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   414..543
                     /label=AAV2 ITR
     intron          1394..1526
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1509..1528
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1571..1589
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1601..1610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1997..2086
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    2135..2256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2146..2165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2200..2219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2418..2547
                     /label=AAV2 ITR
     primer_bind     complement(2566..2583)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2566..2582)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2575..2597)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2724..3179
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3021..3042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3205..3309
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3310..3378,3379..4170)
                     /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      4341..4929
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4830..4849
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
       61 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      361 aggaaacagc tatgaccatg attacgccag atttaattaa ggccttaatt aggctgcgcg
      421 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
      481 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      541 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
      601 tctaacatat cctggtgtgg agtaggggac gctgctctga cagaggctcg ggggcctgag
      661 ctggctctgt gagctgggga ggaggcagac agccaggcct tgtctgcaag cagacctggc
      721 agcattgggc tggccgcccc ccagggcctc ctcttcatgc ccagtgaatg actcaccttg
      781 gcacagacac aatgttcggg gtgggcacag tgcctgcttc ccgccgcacc ccagcccccc
      841 tcaaatgcct tccgagaagc ccattgagca gggggcttgc attgcacccc agcctgacag
      901 cctggcatct tgggataaaa gcagcacagc cccctagggg ctgcccttgc tgtgtggcgc
      961 caccggcggt ggagaacaag gctctattca gcctgtgccc aggaaagggg atcaggggat
     1021 gcccaggcat ggacagtggg tggcaggggg ggagaggagg gctgtctgct tcccagaagt
     1081 ccaaggacac aaatgggtga ggggagagct ctccccatag ctgggctgcg gcccaacccc
     1141 accccctcag gctatgccag ggggtgttgc caggggcacc cgggcatcgc cagtctagcc
     1201 cactccttca taaagccctc gcatcccagg agcgagcaga gccagagcag gttggagagg
     1261 agacgcatca cctccgctgc tcgcaagctt tattgcggta gtttatcaca gttaaattgc
     1321 taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg
     1381 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     1441 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     1501 catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg
     1561 ctagagtact taatacgact cactataggc tagcctcgag gccgccatgg cgcctgtgaa
     1621 aaagcttgtg gcgaaggggg gcaaaaaaaa gaagcaggtt ttgaagttca ccctggactg
     1681 cactcaccct gtagaagatg gaatcatgga tgctgccaat tttgagcagt tcctccagga
     1741 gagaatcaag gtgaatggga aagctggcaa cctcggcgga ggagtcgtga ccatcgaacg
     1801 cagcaagagc aagatcactg tcacttcaga ggtgcctttc tccaaaaggt atttgaaata
     1861 tctcaccaaa aaatatttga agaagaacaa cctccgagac tggctgcgtg ttgtcgccaa
     1921 cagcaaagag agttacgagc tgcgttactt ccagattaac caggatgaag aggaggagga
     1981 agacgaggat gaattctacc catacgatgt tcctgactat gcgggctatc cctatgacgt
     2041 cccggactat gcaggatcct atccatatga cgttccagat tacgctgctc agtagtctag
     2101 agtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2161 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2221 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2281 atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa
     2341 tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg
     2401 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2461 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2521 ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt
     2581 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2641 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2701 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4921 ctatggaaaa acgccagcaa cgcggccttt ttacg
//
