LOCUS       sequence-309124-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated (AA 1-690) murine Thrombospondin-1 (THBS1) 
            protein with HA-tag at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  AUTHORS   Bray ER, Yungher BJ, Levay K, Ribeiro M, Dvoryanchikov G, Ayupe AC, 
            Thakor K, Marks V, Randolph M, Danzi MC, Schmidt TM, Chaudhari N, 
            Lemmon VP, Hattar S, Park KK
  TITLE     Thrombospondin-1 Mediates Axon Regeneration in Retinal Ganglion 
            Cells.
  JOURNAL   Neuron. 2019 Aug 21;103(4):642-657.e7. doi: 
            10.1016/j.neuron.2019.05.044. Epub 2019 Jun 26.
  PUBMED    31255486
REFERENCE   2  (bases 1 to 5708)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2019.05.044"; journalName: "Neuron"; date: 
            "2019-08-21- 21"; volume: "103"; issue: "4"; pages: "642-657.e7"
COMMENT     Sequence Label: pAAV.CMV.SV40.THBS1-deltaTSR3-CTD-HA.SV40(polyA)
FEATURES             Location/Qualifiers
     source          1..5708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   18..147
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        300..503
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     454..474
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     500..524
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2774..2800
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     2857..2876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2988..3128
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2988..3117
                     /label=AAV2 ITR
     rep_origin      3203..3658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3290..3309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3500..3521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3675..3694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3794..3816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3854..3872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3940..4044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4045..4113,4114..4905)
                     /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     complement(4263..4282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5076..5664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5565..5584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccttttgct cacatgtcct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
       61 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      121 tggccaactc catcactagg ggttcctgcg gccgctcgtg acctaggcat atgccaagta
      181 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      241 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      301 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      361 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      421 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      481 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      541 cggactctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta ctgattctaa
      601 ttgtttctct cttttagatt ccaacctttg gaactgaatt ccgcgggccc gggatcctct
      661 agagtcgacc tgcagaagct tcgataagct tatgaccatg gagctcctgc ggggactagg
      721 tgtcctgttc ctgttgcata tgtgtggaag caaccgcatt ccagagtctg ggggagataa
      781 cggtgtgttt gacatctttg aactcattgg aggtgcacga aggggccccg gtcgccgact
      841 ggtgaagggc caagatctat ccagccccgc cttccgcatt gagaatgcca acctgatccc
      901 cgctgtgccg gatgacaagt tccaagacct actggacgct gtgtgggccg acaaaggctt
      961 catcttcctg gcttccttga ggcagatgaa gaagacccgg ggcacactcc tggctgtgga
     1021 acggaaagac aacactggcc agatcttcag tgtggtctcc aacggcaaag ctggcaccct
     1081 ggacctgagc ctgagcctgc cagggaagca acaagtggtg tcagtggagg aagctctcct
     1141 ggccactggc cagtggaaga gcatcacgct gtttgttcaa gaggaccggg ctcaactcta
     1201 catagactgt gataagatgg agagcgcgga gctggatgta cccatccaga gcatcttcac
     1261 cagggatctg gccagcgttg ccaggctccg agttgcaaag ggagatgtca atgacaattt
     1321 tcagggggtg ctgcagaatg tgaggtttgt ctttggaacc accccagaag acattctcag
     1381 gaacaaaggc tgctccagct cagctaccaa cgtccttctt acccttgaca acaacgtggt
     1441 gaacggttcc agccctgcta tccgcaccaa ctacatcggc cacaaaacaa aggacctcca
     1501 agctatctgt ggcctctcct gtgatgaact atccagcatg gtcctggaac tgaagggcct
     1561 gcgcaccatc gtgaccactc tgcaggacag catccgaaaa gtgacggaag agaacagaga
     1621 gctggtcagt gagctgaagc ggcctcccct ctgctttcac aatggagtcc agtacaagaa
     1681 caacgaggag tggactgtag acagttgcac agagtgtcac tgccagaact cggttaccat
     1741 ctgcaaaaag gtgtcctgtc ccatcatgcc ctgctccaac gccacagttc ctgatggtga
     1801 atgctgccca cggtgctggc ccagcgactc tgctgacgat ggctggtctc cctggtctga
     1861 gtggacctcc tgctctgcca catgtggcaa tggaattcag caacgtggtc gttcctgtga
     1921 cagcctcaac aacagatgcg agggctcttc ggtacagaca aggacctgcc acattcagga
     1981 gtgtgacaaa agatttaaac aggatggtgg ctggagtcac tggtctccat ggtcgtcctg
     2041 ttctgtgacc tgtggtgacg gtgtgatcac aaggatccgg ctctgcaact cccccagccc
     2101 ccagatgaac gggaagccct gtgaaggtga agcccgggag accaaagcct gcaagaaaga
     2161 cgcctgccca attaatggag gctggggtcc ctggtcacca tgggacatct gctctgtcac
     2221 ctgtggagga ggagtgcaga gacgcagccg actctgtaac aaccccacac cccagtttgg
     2281 aggcaaagac tgtgttggcg atgtgacaga aaatcaagtt tgcaacaagc aggactgccc
     2341 aattgatgga tgcctgtcca atccctgctt tgctggtgcc aagtgtacta gctaccctga
     2401 tggtagctgg aaatgtggtg cgtgtcctcc tggctacagt ggaaatggca tccagtgcaa
     2461 agacgtcgat gagtgcaaag aagtgcctga tgcttgcttc aatcacaacg gagaacatcg
     2521 gtgcaagaac acagatcctg gctacaactg cctgccctgc ccaccacgat tcactggctc
     2581 acagcccttc ggccgaggtg tcgaacatgc catggccaac aaacaggtgt acaaaccgcg
     2641 aaacccctgc acggacggga cgcatgactg caacaagaac gctaagtgca actacctggg
     2701 tcactacagt gaccccatgt accgctgtga gtgcaagccc ggctatgcag gcaatggcat
     2761 catctgcgca gcttacccat acgatgttcc agattacgct tagctcgaga gatctaggcc
     2821 gctaggcctc acctgcgatc tcgatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2881 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2941 tttcaggttc agggggaggt gtgggaggtt ttttaaactg cggccgcagg aacccctagt
     3001 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3061 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3121 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3181 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3241 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     3541 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3661 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3721 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3781 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3841 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5701 ttttgctg
//
