LOCUS       sequence-500888-        5967 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  Overexpression of C-terminal fragment of mouse Sparcl1 with Myc and 
            6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5967)
  TITLE     Astrocyte-Microglia Crosstalk Through Hevin and Toll-like Receptor 
            Signaling Controls Developmental Thalamocortical Synapse Refinement
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 23, 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            "Unpublished"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-mSLF-Myc-His
FEATURES             Location/Qualifiers
     source          1..5967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..57
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(117..133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(117..133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(130..152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    141..157
                     /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(165..171,172..189,190..195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    210..231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(348..365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(519..1107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(599..618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1278..2069,2070..2138))
                     /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     1901..1920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2139..2243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2322..2341)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2513..2892
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2893..3096
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3047..3067
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3141..3160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3141..3159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3177..3186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3183..3242
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4164..4193
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4209..4226
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4249..4266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4255..4479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4525..4953
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4612..4631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4822..4843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4962..4982)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4967..5296
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5147..5282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5209..5228
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5344..5391
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5410..5784
                     /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"
     polyA_signal    join(5914..5967,1..68)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(5951..5967,1..3))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
       61 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
      121 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
      181 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
      241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
      301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
      361 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      421 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      481 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      541 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      601 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      661 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      721 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      781 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      841 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      901 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      961 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1021 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1081 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1141 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1201 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1261 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1321 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1381 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1441 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1501 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1561 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1621 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1681 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1741 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1801 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1861 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1921 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1981 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2041 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2101 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2161 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2221 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     2281 tcgggagatc gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     2341 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     2401 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     2461 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     2521 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     2581 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2641 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2701 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2761 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2821 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2881 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2941 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     3001 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     3061 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     3121 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcca
     3181 ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg
     3241 gtgacgcggc ccagccggcc gaaaggatgc attccctctc ctattacctc aaatatggcg
     3301 gcggcgagga gacaacgact ggcgagagtg agaaccggag ggaggctgca gacaaccaag
     3361 aggccaagaa agctgagagc tcaccaaatg ctgaaccttc agatgagggc aactcaaggg
     3421 agcacagtgc tggttcttgc acgaacttcc aatgtaaaag gggacacatt tgcaaaaccg
     3481 atccacaagg gaaacctcac tgtgtttgcc aagatccaga gacttgtccc cctgcaaaaa
     3541 tcctagatca ggcttgtggc actgacaacc aaacctacgc cagctcctgt cacctgtttg
     3601 ctaccaagtg caggctggag gggaccaaaa agggacacca actgcagctg gattacttcg
     3661 gagcttgcaa atctattcct gcttgtacgg actttgaagt ggctcagttt cccctgcgga
     3721 tgagagactg gctcaaaaac atcctcatgc agctttatga accaaatccc aaacatggcg
     3781 gctatctcaa tgaaaagcaa agaagcaaag tcaaaaaaat ttacctggat gagaagagac
     3841 tcttggctgg agaccatccc attgaacttc tcttgaggga ctttaagaaa aactaccaca
     3901 tgtatgtgta tcctgtgcac tggcagttta atgaactgga ccagcatcct gcagacagga
     3961 tcttgacaca ctctgaactt gctcctctgc gagcttccct ggtgcccatg gaacactgca
     4021 taactcgctt ctttgaggag tgtgacccca acaaggataa gcacatcacc ttgaaggaat
     4081 ggggccactg ctttggaatt aaagaggagg atatagatga aaacctcctc tttctcgagg
     4141 aagcgctctc tctagaaggg cccgaacaaa aactcatctc agaagaggat ctgaatagcg
     4201 ccgtcgacca tcatcatcat catcattgag tttaaacccg ctgatcagcc tcgactgtgc
     4261 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4321 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4381 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4441 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     4501 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4561 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4621 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4681 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4741 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4801 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4861 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4921 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4981 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5041 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5101 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     5161 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5221 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5281 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     5341 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     5401 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     5461 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     5521 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     5581 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     5641 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     5701 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     5761 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
     5821 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5881 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5941 tacaaataaa gcaatagcat cacaaat
//
