LOCUS       sequence-497595-        5489 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  AAV mediated overexpression of C-terminal fragment of mouse Sparcl1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  TITLE     Astrocyte-Microglia Crosstalk Through Hevin and Toll-like Receptor 
            Signaling Controls Developmental Thalamocortical Synapse Refinement
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5489)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 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            "Unpublished"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-mSLF-Myc-His
FEATURES             Location/Qualifiers
     source          1..5489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    115..136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(151..156,157..174,175..181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    189..205
                     /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     194..216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     213..229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     213..229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   267..396
                     /label=AAV2 ITR
     intron          1247..1379
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1362..1381
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1424..1442
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1446..1455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1452..1511
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2433..2462
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2478..2495
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2522..2643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2533..2552)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2587..2606
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2805..2934
                     /label=AAV2 ITR
     primer_bind     complement(2953..2970)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2953..2969)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2962..2984)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3111..3566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3198..3217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3408..3429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3592..3696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3697..3765,3766..4557)
                     /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      4728..5316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5217..5236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      181 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      241 cagatttaat taaggcctta attaggctgc gcgctcgctc gctcactgag gccgcccggg
      301 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      361 gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat
      421 gctacttatc tacgtagcca tgctctagga agatctaaca tatcctggtg tggagtaggg
      481 gacgctgctc tgacagaggc tcgggggcct gagctggctc tgtgagctgg ggaggaggca
      541 gacagccagg ccttgtctgc aagcagacct ggcagcattg ggctggccgc cccccagggc
      601 ctcctcttca tgcccagtga atgactcacc ttggcacaga cacaatgttc ggggtgggca
      661 cagtgcctgc ttcccgccgc accccagccc ccctcaaatg ccttccgaga agcccattga
      721 gcagggggct tgcattgcac cccagcctga cagcctggca tcttgggata aaagcagcac
      781 agccccctag gggctgccct tgctgtgtgg cgccaccggc ggtggagaac aaggctctat
      841 tcagcctgtg cccaggaaag gggatcaggg gatgcccagg catggacagt gggtggcagg
      901 gggggagagg agggctgtct gcttcccaga agtccaagga cacaaatggg tgaggggaga
      961 gctctcccca tagctgggct gcggcccaac cccaccccct caggctatgc cagggggtgt
     1021 tgccaggggc acccgggcat cgccagtcta gcccactcct tcataaagcc ctcgcatccc
     1081 aggagcgagc agagccagag caggttggag aggagacgca tcacctccgc tgctcgcaag
     1141 ctttattgcg gtagtttatc acagttaaat tgctaacgca gtcagtgctt ctgacacaac
     1201 agtctcgaac ttaagctgca gaagttggtc gtgaggcact gggcaggtaa gtatcaaggt
     1261 tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt
     1321 gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg
     1381 tgtccactcc cagttcaatt acagctctta aggctagagt acttaatacg actcactata
     1441 ggctagccac catggagaca gacacactcc tgctatgggt actgctgctc tgggttccag
     1501 gttccactgg tgacgcggcc cagccggccg aaaggatgca ttccctctcc tattacctca
     1561 aatatggcgg cggcgaggag acaacgactg gcgagagtga gaaccggagg gaggctgcag
     1621 acaaccaaga ggccaagaaa gctgagagct caccaaatgc tgaaccttca gatgagggca
     1681 actcaaggga gcacagtgct ggttcttgca cgaacttcca atgtaaaagg ggacacattt
     1741 gcaaaaccga tccacaaggg aaacctcact gtgtttgcca agatccagag acttgtcccc
     1801 ctgcaaaaat cctagatcag gcttgtggca ctgacaacca aacctacgcc agctcctgtc
     1861 acctgtttgc taccaagtgc aggctggagg ggaccaaaaa gggacaccaa ctgcagctgg
     1921 attacttcgg agcttgcaaa tctattcctg cttgtacgga ctttgaagtg gctcagtttc
     1981 ccctgcggat gagagactgg ctcaaaaaca tcctcatgca gctttatgaa ccaaatccca
     2041 aacatggcgg ctatctcaat gaaaagcaaa gaagcaaagt caaaaaaatt tacctggatg
     2101 agaagagact cttggctgga gaccatccca ttgaacttct cttgagggac tttaagaaaa
     2161 actaccacat gtatgtgtat cctgtgcact ggcagtttaa tgaactggac cagcatcctg
     2221 cagacaggat cttgacacac tctgaacttg ctcctctgcg agcttccctg gtgcccatgg
     2281 aacactgcat aactcgcttc tttgaggagt gtgaccccaa caaggataag cacatcacct
     2341 tgaaggaatg gggccactgc tttggaatta aagaggagga tatagatgaa aacctcctct
     2401 ttctcgagga agcgctctct ctagaagggc ccgaacaaaa actcatctca gaagaggatc
     2461 tgaatagcgc cgtcgaccat catcatcatc atcattgagc ggccgcttcg agcagacatg
     2521 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2581 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2641 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2701 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     2761 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     2821 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     2881 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagccttaa
     2941 ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     3001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     3061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     3121 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3181 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3241 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3301 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3361 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3421 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3481 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     3541 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     3601 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3661 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3721 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3781 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3841 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3901 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3961 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4021 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4081 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4141 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4201 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4261 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4321 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4381 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4441 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4501 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4561 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4621 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4681 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4741 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4801 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4861 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4921 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4981 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5041 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5101 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5161 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5221 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5281 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5341 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5401 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     5461 accgagcgca gcgagtcagt gagcgagga
//
