LOCUS       sequence-497522-        6798 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  Overexpression of mouse Sparcl1 with deletion of a.a. 330-550 with 
            Myc and 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6798)
  TITLE     Astrocyte-Microglia Crosstalk Through Hevin and Toll-like Receptor 
            Signaling Controls Developmental Thalamocortical Synapse Refinement
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6798)
  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            "Unpublished"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: pAP-Tag5-IgKsp-mHevin∆DE-Myc-His
FEATURES             Location/Qualifiers
     source          1..6798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    142..263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(179..198)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     233..252
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(312..328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(312..328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(325..347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     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)."
     promoter        complement(join(360..366,367..384,385..390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    405..426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(543..560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(714..1302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(794..813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1473..2264,2265..2333))
                     /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"
     primer_bind     2096..2115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2334..2438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2517..2536)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2708..3087
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3088..3291
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3242..3262
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3336..3355
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3336..3354
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3372..3381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3378..3437
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5190..5219
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5235..5252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5275..5292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5281..5505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5551..5979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5638..5657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5848..5869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5988..6008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5993..6322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6173..6308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6235..6254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6370..6417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(6436..6798,1..12)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
       61 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
      121 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
      181 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      241 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
      301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1441 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1501 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1561 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1621 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1681 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1741 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1801 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1861 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1921 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1981 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2041 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2101 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2161 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2221 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2281 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2341 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2401 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2461 cctgacgtcg acggatcggg agatcgatct cccgatcccc tatggtgcac tctcagtaca
     2521 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     2581 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     2641 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     2701 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     2761 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2821 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2881 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2941 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     3001 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     3061 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     3121 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3181 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3241 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     3301 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     3361 caagctggct agccaccatg gagacagaca cactcctgct atgggtactg ctgctctggg
     3421 ttccaggttc cactggtgac gcggcccagc cggccatccc gacaagtaca aggtttctct
     3481 ctgaccactc caacccaact actgcaacac tggtgacacc ggaagacgct acagtcccca
     3541 ttgccggggt tgaagctaca gcagacatag aaaaccatcc caatgacaag gctgaaaaac
     3601 cttcagcact taattcagaa gaggaaactc atgaacagtc aacagagcag gacaaaacct
     3661 acagcttcga ggtggacctg aaggatgagg aggatggaga tggggattta agtgtagatc
     3721 caacggaagg aacactaaca ctggatctac aagaaggcac aagtgagcct caacagaaaa
     3781 gtctcccgga gaacggcgat ttccccgcga ccgtgtccac ttcctatgtg gatcctaacc
     3841 aacgcgcaaa catcacaaag ggaaaggaga gtcaggagca acctgtaagt gactcacacc
     3901 agcaaccgaa tgaaagcagc aagcaaaccc aagacttaaa ggctgaagaa agccagacac
     3961 aagatccaga cattcccaat gaagaagagg aagaagaaga ggacgaagaa gaggaagaag
     4021 aggaagagcc ggaagacatt ggtgccccca gtgataacca agaggaggga aaagaacctc
     4081 tggaggagca gcctaccagc aagtgggaag gaaacagaga gcaatctgat gacaccttag
     4141 aagagtccag tcagcccact cagataagca agacagagaa gcatcaatct gagcaaggaa
     4201 accaagggca ggagagtgac tctgaggcag aaggagagga caaggctgca ggcagcaagg
     4261 aacacattcc acatacagag cagcaggacc aagaagggaa agctggcctt gaagctattg
     4321 gcaaccagaa ggacactgat gagaaggccg tttccacaga acctaccgat gctgccgtgg
     4381 tgcctaggag tcacggagga gctggtgata acgggggcgg ggatgactct aagcatggtg
     4441 caggcgatga ctacttcggt tcttgcacga acttccaatg taaaagggga cacatttgca
     4501 aaaccgatcc acaagggaaa cctcactgtg tttgccaaga tccagagact tgtccccctg
     4561 caaaaatcct agatcaggct tgtggcactg acaaccaaac ctacgccagc tcctgtcacc
     4621 tgtttgctac caagtgcagg ctggagggga ccaaaaaggg acaccaactg cagctggatt
     4681 acttcggagc ttgcaaatct attcctgctt gtacggactt tgaagtggct cagtttcccc
     4741 tgcggatgag agactggctc aaaaacatcc tcatgcagct ttatgaacca aatcccaaac
     4801 atggcggcta tctcaatgaa aagcaaagaa gcaaagtcaa aaaaatttac ctggatgaga
     4861 agagactctt ggctggagac catcccattg aacttctctt gagggacttt aagaaaaact
     4921 accacatgta tgtgtatcct gtgcactggc agtttaatga actggaccag catcctgcag
     4981 acaggatctt gacacactct gaacttgctc ctctgcgagc ttccctggtg cccatggaac
     5041 actgcataac tcgcttcttt gaggagtgtg accccaacaa ggataagcac atcaccttga
     5101 aggaatgggg ccactgcttt ggaattaaag aggaggatat agatgaaaac ctcctctttc
     5161 tcgaggaagc gctctctcta gaagggcccg aacaaaaact catctcagaa gaggatctga
     5221 atagcgccgt cgaccatcat catcatcatc attgagttta aacccgctga tcagcctcga
     5281 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5341 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5401 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5461 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     5521 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     5581 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5641 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5701 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5761 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5821 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5881 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     5941 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     6001 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6061 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6121 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6181 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6241 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     6301 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     6361 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     6421 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     6481 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     6541 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     6601 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     6661 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     6721 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     6781 gtgcacttcg tggccgag
//
