LOCUS       sequence-495627-        7044 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Overexpression of full length mouse Sparcl1 with Myc and 6xHis tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7044)
  TITLE     Astrocyte-Microglia Crosstalk Through Hevin and Toll-like Receptor 
            Signaling Controls Developmental Thalamocortical Synapse Refinement
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 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            "Unpublished"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-Myc-His
FEATURES             Location/Qualifiers
     source          1..7044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..30
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(249..353)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(432..451)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        623..1002
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1003..1206
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1157..1177
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1251..1270
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1251..1269
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1287..1296
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1293..1352
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             3351..3380
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3396..3413
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3436..3453)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3442..3666
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3712..4140
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3799..3818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4009..4030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4149..4169)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4154..4483
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4334..4469
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4396..4415
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4531..4578
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4597..4971
                     /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    5101..5222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5138..5157)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5192..5211
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5271..5287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5271..5287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5284..5306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5295..5311
                     /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(5319..5325,5326..5343,5344..5349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5364..5385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5502..5519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5673..6261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5753..5772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6432..7044,1..179,180..248))
                     /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 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
       61 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
      121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      361 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctatgg
      421 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      481 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      541 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      601 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      661 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      721 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      781 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      841 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      901 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      961 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     1021 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     1081 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1141 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1201 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     1261 cactataggg agacccaagc tggctagcca ccatggagac agacacactc ctgctatggg
     1321 tactgctgct ctgggttcca ggttccactg gtgacgcggc ccagccggcc aggcgcgcgc
     1381 gccgtacgta cgaagcttac gtaagatctg ccaccatgat cccgacaagt acaaggtttc
     1441 tctctgacca ctccaaccca actactgcaa cactggtgac accggaagac gctacagtcc
     1501 ccattgccgg ggttgaagct acagcagaca tagaaaacca tcccaatgac aaggctgaaa
     1561 aaccttcagc acttaattca gaagaggaaa ctcatgaaca gtcaacagag caggacaaaa
     1621 cctacagctt cgaggtggac ctgaaggatg aggaggatgg agatggggat ttaagtgtag
     1681 atccaacgga aggaacacta acactggatc tacaagaagg tacaagtgag cctcaacaga
     1741 aaagtctccc ggaaaacggc gatttccccg cgaccgtgtc cacttcctat gtggatccta
     1801 accaacgcgc aaacatcaca aagggaaagg agagtcagga gcaacctgta agtgactcac
     1861 accagcaacc gaatgaaagc agcaagcaaa cccaagactt aaaggctgaa gaaagccaga
     1921 cacaagatcc agacattccc aatgaagaag aggaagaaga agaggaagaa gaagaggaag
     1981 aagaggaaga gccggaagac attggtgccc ccagtgataa ccaagaggag ggaaaagaac
     2041 ctctggagga gcagcctacc agcaagtggg aaggaaacag agagcaatct gatgacacct
     2101 tagaagagtc cagtcagccc actcagataa gcaagacaga gaagcatcaa tctgagcaag
     2161 gaaaccaagg gcaggagagt gactctgagg cagaaggaga ggacaaggct gcaggcagca
     2221 aggaacacat tccacataca gagcagcagg accaagaagg gaaagctggc cttgaagcta
     2281 ttggcaacca gaaggacact gatgagaagg ccgtttccac agaacctacc gatgctgccg
     2341 tggtgcctag gagtcacgga ggagctggtg ataacggggg cggggatgac tctaagcatg
     2401 gtgcaggcga tgactacttc atccccagcc aggaattcct agaggccgaa aggatgcatt
     2461 ccctctccta ttacctcaaa tatggcggcg gcgaggagac aacgactggc gagagtgaga
     2521 accggaggga ggctgcagac aaccaagagg ccaagaaagc tgagagctca ccaaatgctg
     2581 aaccttcaga tgagggcaac tcaagggagc acagtgctgg ttcttgcacg aacttccaat
     2641 gtaaaagggg acacatttgc aaaaccgatc cacaagggaa acctcactgt gtttgccaag
     2701 atccagagac ttgtccccct gcaaaaatcc tagatcaggc ttgtggcact gacaaccaaa
     2761 cctacgccag ctcctgtcac ctgtttgcta ccaagtgcag gctggagggg accaaaaagg
     2821 gacaccaact gcagctggat tacttcggag cttgcaaatc tattcctgct tgtacggact
     2881 ttgaagtggc tcagtttccc ctgcggatga gagactggct caaaaacatc ctcatgcagc
     2941 tttatgaacc aaatcccaaa catggcggct atctcaatga aaagcaaaga agcaaagtca
     3001 aaaaaattta cctggatgag aagagactct tggctggaga ccatcccatt gaacttctct
     3061 tgagggactt taagaaaaac taccacatgt atgtgtatcc tgtgcactgg cagtttaatg
     3121 aactggacca gcatcctgca gacaggatct tgacacactc tgaacttgct cctctgcgag
     3181 cttccctggt gcccatggaa cactgcataa ctcgcttctt tgaggagtgt gaccccaaca
     3241 aggataagca catcaccttg aaggaatggg gccactgctt tggaattaaa gaggaggata
     3301 tagatgaaaa cctcctcttt ctcgaggaag cgctctctct agaagggccc gaacaaaaac
     3361 tcatctcaga agaggatctg aatagcgccg tcgaccatca tcatcatcat cattgagttt
     3421 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3481 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3541 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3601 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3661 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     3721 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3781 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     3841 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     3901 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     3961 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4021 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     4081 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4141 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4201 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4261 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4321 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4381 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4441 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4501 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     4561 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
     4621 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     4681 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     4741 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     4801 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     4861 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     4921 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
     4981 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5041 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5101 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5161 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5221 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7021 cgaccgagtt gctcttgccc ggcg
//
