LOCUS       sequence-309333-        6499 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses truncated (AA 1-954) 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 6499)
  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 6499)
  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-deltaCTD-HA.SV40(polyA)
FEATURES             Location/Qualifiers
     source          1..6499
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        95..298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     249..269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     295..319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3360..3386
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3443..3462
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3574..3714
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3574..3703
                     /label=AAV2 ITR
     rep_origin      3789..4244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4086..4107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4261..4280)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4380..4402
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4440..4458)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4526..4630
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4631..4699,4700..5491)
                     /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(4849..4868)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5662..6250
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6151..6170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6312..6441
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
       61 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
      301 tttagtgaac cgtcagatcg cctggagacg ccatccggac tctaaggtaa atataaaatt
      361 tttaagtgta taatgtgtta aactactgat tctaattgtt tctctctttt agattccaac
      421 ctttggaact gaattccgcg ggcccgggat cctctagagt cgacctgcag aagcttcata
      481 agcttatgac catggagctc ctgcggggac taggtgtcct gttcctgttg catatgtgtg
      541 gaagcaaccg cattccagag tctgggggag ataacggtgt gtttgacatc tttgaactca
      601 ttggaggtgc acgaaggggc cccggtcgcc gactggtgaa gggccaagat ctatccagcc
      661 ccgccttccg cattgagaat gccaacctga tccccgctgt gccggatgac aagttccaag
      721 acctactgga cgctgtgtgg gccgacaaag gcttcatctt cctggcttcc ttgaggcaga
      781 tgaagaagac ccggggcaca ctcctggctg tggaacggaa agacaacact ggccagatct
      841 tcagtgtggt ctccaacggc aaagctggca ccctggacct gagcctgagc ctgccaggga
      901 agcaacaagt ggtgtcagtg gaggaagctc tcctggccac tggccagtgg aagagcatca
      961 cgctgtttgt tcaagaggac cgggctcaac tctacataga ctgtgataag atggagagcg
     1021 cggagctgga tgtacccatc cagagcatct tcaccaggga tctggccagc gttgccaggc
     1081 tccgagttgc aaagggagat gtcaatgaca attttcaggg ggtgctgcag aatgtgaggt
     1141 ttgtctttgg aaccacccca gaagacattc tcaggaacaa aggctgctcc agctcagcta
     1201 ccaacgtcct tcttaccctt gacaacaacg tggtgaacgg ttccagccct gctatccgca
     1261 ccaactacat cggccacaaa acaaaggacc tccaagctat ctgtggcctc tcctgtgatg
     1321 aactatccag catggtcctg gaactgaagg gcctgcgcac catcgtgacc actctgcagg
     1381 acagcatccg aaaagtgacg gaagagaaca gagagctggt cagtgagctg aagcggcctc
     1441 ccctctgctt tcacaatgga gtccagtaca agaacaacga ggagtggact gtagacagtt
     1501 gcacagagtg tcactgccag aactcggtta ccatctgcaa aaaggtgtcc tgtcccatca
     1561 tgccctgctc caacgccaca gttcctgatg gtgaatgctg cccacggtgc tggcccagcg
     1621 actctgctga cgatggctgg tctccctggt ctgagtggac ctcctgctct gccacatgtg
     1681 gcaatggaat tcagcaacgt ggtcgttcct gtgacagcct caacaacaga tgcgagggct
     1741 cttcggtaca gacaaggacc tgccacattc aggagtgtga caaaagattt aaacaggatg
     1801 gtggctggag tcactggtct ccatggtcgt cctgttctgt gacctgtggt gacggtgtga
     1861 tcacaaggat ccggctctgc aactccccca gcccccagat gaacgggaag ccctgtgaag
     1921 gtgaagcccg ggagaccaaa gcctgcaaga aagacgcctg cccaattaat ggaggctggg
     1981 gtccctggtc accatgggac atctgctctg tcacctgtgg aggaggagtg cagagacgca
     2041 gccgactctg taacaacccc acaccccagt ttggaggcaa agactgtgtt ggcgatgtga
     2101 cagaaaatca agtttgcaac aagcaggact gcccaattga tggatgcctg tccaatccct
     2161 gctttgctgg tgccaagtgt actagctacc ctgatggtag ctggaaatgt ggtgcgtgtc
     2221 ctcctggcta cagtggaaat ggcatccagt gcaaagacgt cgatgagtgc aaagaagtgc
     2281 ctgatgcttg cttcaatcac aacggagaac atcggtgcaa gaacacagat cctggctaca
     2341 actgcctgcc ctgcccacca cgattcactg gctcacagcc cttcggccga ggtgtcgaac
     2401 atgccatggc caacaaacag gtgtgcaaac cgcgaaaccc ctgcacggac gggacgcatg
     2461 actgcaacaa gaacgctaag tgcaactacc tgggtcacta cagtgacccc atgtaccgct
     2521 gtgagtgcaa gcccggctat gcaggcaatg gcatcatctg cggagaggac acagacctgg
     2581 acggctggcc taatgaaaac ctggtgtgtg tggccaacgc aacctaccac tgcaaaaagg
     2641 acaactgccc caaccttccc aactcggggc aggaagacta tgacaaggac gggattggcg
     2701 atgcctgcga tgatgacgat gacaacgaca agatccccga tgacagggac aactgtccat
     2761 tccattacaa cccagcccag tatgactatg acagagatga tgtgggagac cgctgtgaca
     2821 actgccccta caaccacaac cctgaccaag cagacacaga caaaaacggg gagggcgatg
     2881 cctgtgctgt ggacatcgat ggagatggaa tcctcaatga acgagacaac tgccagtacg
     2941 tttacaacgt ggaccagagg gacacggaca tggatggggt tggagatcag tgtgacaact
     3001 gccccctgga acacaatcca gaccagctgg actctgactc agacctcata ggggacactt
     3061 gtgacaacaa tcaggacatc gatgaggatg gccatcagaa caatctggac aactgtccct
     3121 atgtgcctaa tgccaaccag gccgaccatg ataaagatgg caaaggagat gcctgtgacc
     3181 atgacgatga caatgacggc atccctgatg acagagacaa ctgcaggctg gtgcccaatc
     3241 ctgaccagaa ggactctgat ggtgatggcc gaggtgacgc ctgcaaagac gactttgacc
     3301 atgacaatgt gccagatatt gatgacatct gtcctgagaa ttttgacatc agtgcagctt
     3361 acccatacga tgttccagat tacgcttagc tcgagagatc taggccgcta ggcctcacct
     3421 gcgatctcga tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3481 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3541 ggaggtgtgg gaggtttttt aaactgcggc cgcaggaacc cctagtgatg gagttggcca
     3601 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3661 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3721 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3781 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3841 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3901 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3961 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4021 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4081 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4141 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4201 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4261 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4321 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4381 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4441 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4621 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4681 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4741 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4801 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4861 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4921 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4981 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5041 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5101 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5161 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5221 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5281 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5341 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5401 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5461 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5521 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5581 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5641 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5701 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5761 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5821 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5881 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5941 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6001 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6061 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6121 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6181 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6241 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6301 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     6361 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     6421 actccatcac taggggttcc tgcggccgct cgtgacctag gcatatgcca agtacgcccc
     6481 ctattgacgt caatgacgg
//
