LOCUS       sequence-309129-        6637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Thrombospondin-1 (THBS1) with deletion (AA 383-548)
            and HA-tag at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6637)
  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 6637)
  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-deltaTSR1-HA.SV40(polyA)
FEATURES             Location/Qualifiers
     source          1..6637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..262
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     213..233
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     259..283
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3462..3488
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3545..3564
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3676..3816
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3676..3805
                     /label=AAV2 ITR
     rep_origin      3891..4346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3978..3997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4188..4209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4363..4382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4482..4504
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4542..4560)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4628..4732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4733..4801,4802..5593)
                     /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(4951..4970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5764..6352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6253..6272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6414..6543
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
       61 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      121 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      181 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      241 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
      301 ggactctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
      361 tgtttctctc ttttagattc caacctttgg aactgaattc cgcgggcccg ggatcctcta
      421 gagtcgacct gcagaagctt atgaccatgg agctcctgcg gggactaggt gtcctgttcc
      481 tgttgcatat gtgtggaagc aaccgcattc cagagtctgg gggagataac ggtgtgtttg
      541 acatctttga actcattgga ggtgcacgaa ggggccccgg tcgccgactg gtgaagggcc
      601 aagatctatc cagccccgcc ttccgcattg agaatgccaa cctgatcccc gctgtgccgg
      661 atgacaagtt ccaagaccta ctggacgctg tgtgggccga caaaggcttc atcttcctgg
      721 cttccttgag gcagatgaag aagacccggg gcacactcct ggctgtggaa cggaaagaca
      781 acactggcca gatcttcagt gtggtctcca acggcaaagc tggcaccctg gacctgagcc
      841 tgagcctgcc agggaagcaa caagtggtgt cagtggagga agctctcctg gccactggcc
      901 agtggaagag catcacgctg tttgttcaag aggaccgggc tcaactctac atagactgtg
      961 ataagatgga gagcgcggag ctggatgtac ccatccagag catcttcacc agggatctgg
     1021 ccagcgttgc caggctccga gttgcaaagg gagatgtcaa tgacaatttt cagggggtgc
     1081 tgcagaatgt gaggtttgtc tttggaacca ccccagaaga cattctcagg aacaaaggct
     1141 gctccagctc agctaccaac gtccttctta cccttgacaa caacgtggtg aacggttcca
     1201 gccctgctat ccgcaccaac tacatcggcc acaaaacaaa ggacctccaa gctatctgtg
     1261 gcctctcctg tgatgaacta tccagcatgg tcctggaact gaagggcctg cgcaccatcg
     1321 tgaccactct gcaggacagc atccgaaaag tgacggaaga gaacagagag ctggtcagtg
     1381 agctgaagcg gcctcccctc tgctttcaca atggagtcca gtacaagaac aacgaggagt
     1441 ggactgtaga cagttgcaca gagtgtcact gccagaactc ggttaccatc tgcaaaaagg
     1501 tgtcctgtcc catcatgccc tgctccaacg ccacagttcc tgatggtgaa tgctgcccac
     1561 ggtgctggcc cagcgactct gctgacgatg gcattgatgg atgcctgtcc aatccctgct
     1621 ttgctggtgc caagtgtact agctaccctg atggtagctg gaaatgtggt gcgtgtcctc
     1681 ctggctacag tggaaatggc atccagtgca aagacgtcga tgagtgcaaa gaagtgcctg
     1741 atgcttgctt caatcacaac ggagaacatc ggtgcaagaa cacagatcct ggctacaact
     1801 gcctgccctg cccaccacga ttcactggct cacagccctt cggccgaggt gtcgaacatg
     1861 ccatggccaa caaacaggtg tgcaaaccgc gaaacccctg cacggacggg acgcatgact
     1921 gcaacaagaa cgctaagtgc aactacctgg gtcactacag tgaccccatg taccgctgtg
     1981 agtgcaagcc cggctatgca ggcaatggca tcatctgcgg agaggacaca gacctggacg
     2041 gctggcctaa tgaaaacctg gtgtgtgtgg ccaacgcaac ctaccactgc aaaaaggaca
     2101 actgccccaa ccttcccaac tcggggcagg aagactatga caaggacggg attggcgatg
     2161 cctgcgatga tgacgatgac aacgacaaga tccccgatga cagggacaac tgtccattcc
     2221 attacaaccc agcccagtat gactatgaca gagatgatgt gggagaccgc tgtgacaact
     2281 gcccctacaa ccacaaccct gaccaagcag acacagacaa aaacggggag ggcgatgcct
     2341 gtgctgtgga catcgatgga gatggaatcc tcaatgaacg agacaactgc cagtacgttt
     2401 acaacgtgga ccagagggac acggacatgg atggggttgg agatcagtgt gacaactgcc
     2461 ccctggaaca caatccagac cagctggact ctgactcaga cctcataggg gacacttgtg
     2521 acaacaatca ggacatcgat gaggatggcc atcagaacaa tctggacaac tgtccctatg
     2581 tgcctaatgc caaccaggcc gaccatgata aagatggcaa aggagatgcc tgtgaccatg
     2641 acgatgacaa tgacggcatc cctgatgaca gagacaactg caggctggtg cccaatcctg
     2701 accagaagga ctctgatggt gatggccgag gtgacgcctg caaagacgac tttgaccatg
     2761 acaatgtgcc agatattgat gacatctgtc ctgagaattt tgacatcagt gaaaccgatt
     2821 tccgacaatt ccagatgatt cctctagatc ccaaaggaac ctcccaaaat gaccctaact
     2881 gggttgtccg ccatcagggc aaagaacttg tccagactgt aaactgtgac cctggacttg
     2941 ctgtaggtta tgatgagttt aatgctgtgg acttcagcgg taccttcttc atcaacaccg
     3001 agagagatga tgactacgct ggctttgttt tcggctacca gtccagcagc cgcttctacg
     3061 ttgtgatgtg gaaacaagtc acccagtcct actgggacac caaccccaca agggctcagg
     3121 gatactcagg cctgtctgta aaggttgtga actccaccac cggccctggc gagcacctgc
     3181 ggaatgcact gtggcacaca ggaaacaccc ctggccaggt gcgcaccctg tggcatgacc
     3241 ctcgccacat cggctggaaa gatttcactg catacagatg gcgtctcagc cacaggccaa
     3301 agaccggtta tatcagagtg gtgatgtatg aaggaaagaa aatcatggct gactcgggac
     3361 ccatctatga caaaacctac gccggcggta gactaggcct gttcgtcttc tctcaggaaa
     3421 tggtgttctt ctcagacatg aaatacgagt gtcgagattc ctacccatac gatgttccag
     3481 attacgctta actcgagaga tctaggccgc taggcctcac ctgcgatctc gatgctttat
     3541 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3601 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     3661 ttaaactgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3721 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3781 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     3841 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     3901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3961 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4081 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4201 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4261 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4321 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4381 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4441 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4501 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4561 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
     6421 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
     6481 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     6541 cctgcggccg ctcgtgacct aggcatatgc caagtacgcc ccctattgac gtcaatgacg
     6601 gtaaatggcc cgcctggcat tatgcccagt acatgac
//
