LOCUS Exported 6499 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses truncated (AA 1-954) murine Thrombospondin-1 (THBS1) protein with HA-tag at C-terminus. ACCESSION . VERSION . KEYWORDS pAAV.CMV.SV40.THBS1-deltaCTD-HA.SV40(polyA) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6499 /organism="synthetic DNA construct" /mol_type="other DNA" 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 4631..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 //