LOCUS Exported 5708 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses truncated (AA 1-690) murine Thrombospondin-1 (THBS1) protein with HA-tag at C-terminus. ACCESSION . VERSION . KEYWORDS pAAV.CMV.SV40.THBS1-deltaTSR3-CTD-HA.SV40(polyA) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5708 /organism="synthetic DNA construct" /mol_type="other DNA" 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 4045..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 //