LOCUS       sequence-497579-        6496 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  AAV mediated overexpression of full length mouse Sparcl1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  TITLE     Astrocyte-Microglia Crosstalk Through Hevin and Toll-like Receptor 
            Signaling Controls Developmental Thalamocortical Synapse Refinement
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6496)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-GfaABC1D-mHevin-Myc-His
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      93..681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     582..601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    969..990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1005..1010,1011..1028,1029..1035)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1043..1059
                     /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     1048..1070
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1067..1083
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1067..1083
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     intron          2079..2211
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2194..2213
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        2256..2274
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2278..2287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     2284..2343
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4294..4323
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4339..4356
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4383..4504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4394..4413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4448..4467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4666..4795
                     /label=AAV2 ITR
     primer_bind     complement(4814..4831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4814..4830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4823..4845)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4972..5427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5059..5078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5269..5290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5453..5557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5558..5626,5627..6418)
                     /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"
ORIGIN
        1 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
       61 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      121 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      181 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      241 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      301 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      361 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      421 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      481 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      541 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      601 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      661 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
      721 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
      781 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
      841 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
      901 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      961 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     1021 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     1081 gaccatgatt acgccagatt taattaaggc cttaattagg ctgcgcgctc gctcgctcac
     1141 tgaggccgcc cgggcaaagc ccgggcggcc tcagtgagcg agcgagcgcg cagagaggga
     1201 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
     1261 tctacgtagc catgctctag gaagatctaa catatcctgg tgtggagtag gggacgctgc
     1321 tctgacagag gctcgggggc ctgagctggc tctgtgagct ggggaggagg cagacagcca
     1381 ggccttgtct gcaagcagac ctggcagcat tgggctggcc gccccccagg gcctcctctt
     1441 catgcccagt gaatgactca ccttggcaca gacacaatgt tcggggtggg cacagtgcct
     1501 gcttcccgcc gcaccccagc ccccctcaaa tgccttccga gaagcccatt gagcaggggg
     1561 cttgcattgc accccagcct gacagcctgg catcttggga taaaagcagc acagccccct
     1621 aggggctgcc cttgctgtgt ggcgccaccg gcggtggaga acaaggctct attcagcctg
     1681 tgcccaggaa aggggatcag gggatgccca ggcatggaca gtgggtggca gggggggaga
     1741 ggagggctgt ctgcttccca gaagtccaag gacacaaatg ggtgagggga gagctctccc
     1801 catagctggg ctgcggccca accccacccc ctcaggctat gccagggggt gttgccaggg
     1861 gcacccgggc atcgccagtc tagcccactc cttcataaag ccctcgcatc ccaggagcga
     1921 gcagagccag agcaggttgg agaggagacg catcacctcc gctgctcgca agctttattg
     1981 cggtagttta tcacagttaa attgctaacg cagtcagtgc ttctgacaca acagtctcga
     2041 acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag gttacaagac
     2101 aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct
     2161 gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca ggtgtccact
     2221 cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta taggctagcc
     2281 accatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact
     2341 ggtgacgcgg cccagccggc catcccgaca agtacaaggt ttctctctga ccactccaac
     2401 ccaactactg caacactggt gacaccggaa gacgctacag tccccattgc cggggttgaa
     2461 gctacagcag acatagaaaa ccatcccaat gacaaggctg aaaaaccttc agcacttaat
     2521 tcagaagagg aaactcatga acagtcaaca gagcaggaca aaacctacag cttcgaggtg
     2581 gacctgaagg atgaggagga tggagatggg gatttaagtg tagatccaac ggaaggaaca
     2641 ctaacactgg atctacaaga aggcacaagt gagcctcaac agaaaagtct cccggagaac
     2701 ggcgatttcc ccgcgaccgt gtccacttcc tatgtggatc ctaaccaacg cgcaaacatc
     2761 acaaagggaa aggagagtca ggagcaacct gtaagtgact cacaccagca accgaatgaa
     2821 agcagcaagc aaacccaaga cttaaaggct gaagaaagcc agacacaaga tccagacatt
     2881 cccaatgaag aagaggaaga agaagaggac gaagaagagg aagaagagga agagccggaa
     2941 gacattggtg cccccagtga taaccaagag gagggaaaag aacctctgga ggagcagcct
     3001 accagcaagt gggaaggaaa cagagagcaa tctgatgaca ccttagaaga gtccagtcag
     3061 cccactcaga taagcaagac agagaagcat caatctgagc aaggaaacca agggcaggag
     3121 agtgactctg aggcagaagg agaggacaag gctgcaggca gcaaggaaca cattccacat
     3181 acagagcagc aggaccaaga agggaaagct ggccttgaag ctattggcaa ccagaaggac
     3241 actgatgaga aggccgtttc cacagaacct accgatgctg ccgtggtgcc taggagtcac
     3301 ggaggagctg gtgataacgg gggcggggat gactctaagc atggtgcagg cgatgactac
     3361 ttcatcccca gccaggaatt cctagaggcc gaaaggatgc attccctctc ctattacctc
     3421 aaatatggcg gcggcgagga gacaacgact ggcgagagtg agaaccggag ggaggctgca
     3481 gacaaccaag aggccaagaa agctgagagc tcaccaaatg ctgaaccttc agatgagggc
     3541 aactcaaggg agcacagtgc tggttcttgc acgaacttcc aatgtaaaag gggacacatt
     3601 tgcaaaaccg atccacaagg gaaacctcac tgtgtttgcc aagatccaga gacttgtccc
     3661 cctgcaaaaa tcctagatca ggcttgtggc actgacaacc aaacctacgc cagctcctgt
     3721 cacctgtttg ctaccaagtg caggctggag gggaccaaaa agggacacca actgcagctg
     3781 gattacttcg gagcttgcaa atctattcct gcttgtacgg actttgaagt ggctcagttt
     3841 cccctgcgga tgagagactg gctcaaaaac atcctcatgc agctttatga accaaatccc
     3901 aaacatggcg gctatctcaa tgaaaagcaa agaagcaaag tcaaaaaaat ttacctggat
     3961 gagaagagac tcttggctgg agaccatccc attgaacttc tcttgaggga ctttaagaaa
     4021 aactaccaca tgtatgtgta tcctgtgcac tggcagttta atgaactgga ccagcatcct
     4081 gcagacagga tcttgacaca ctctgaactt gctcctctgc gagcttccct ggtgcccatg
     4141 gaacactgca taactcgctt ctttgaggag tgtgacccca acaaggataa gcacatcacc
     4201 ttgaaggaat ggggccactg ctttggaatt aaagaggagg atatagatga aaacctcctc
     4261 tttctcgagg aagcgctctc tctagaaggg cccgaacaaa aactcatctc agaagaggat
     4321 ctgaatagcg ccgtcgacca tcatcatcat catcattgag cggccgcttc gagcagacat
     4381 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     4441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     4501 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     4561 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc
     4621 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     4681 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4741 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta
     4801 attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4861 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4921 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     4981 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5041 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5101 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5161 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5221 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5281 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5341 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5401 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     5461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5521 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5581 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5641 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5701 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5761 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5821 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5881 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5941 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6001 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6061 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6121 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6181 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6241 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6301 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6361 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6421 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6481 aaggatctag gtgaag
//
