LOCUS Exported 6090 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses murine Vitronectin (VTN) protein with HA-tag at C-terminus. ACCESSION . VERSION . KEYWORDS pAAV.CMV.VTN.C-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6090) 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 6090) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6090 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 49..189 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 60..189 /label=AAV2 ITR enhancer 265..568 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 569..772 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 723..743 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 769..793 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2848..2874 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2885..3361 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3084..3103) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3401..3541 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3401..3530 /label=AAV2 ITR rep_origin 3616..4071 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3703..3722) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3913..3934 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4088..4107) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4207..4229 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4267..4285) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4353..4457 /gene="bla" /label=AmpR promoter CDS 4458..5318 /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(4676..4695) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5489..6077 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5978..5997 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc 61 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg 121 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta 181 ggggttcctg cggccgcacg cgtctagtta ttaatagtaa tcaattacgg ggtcattagt 241 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 301 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 361 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 421 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 481 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 541 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 601 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 661 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 721 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt 781 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg 841 ggaccgatcc agcctccgcg gattcgaatc ccggccggga acggtgcatt ggaacgcgga 901 ttccccgtgc caagagtgac gtaagtaccg cctatagagt ctataggccc acaaaaaatg 961 ctttcttctt ttaatatact tttttgttta tcttatttct aatactttcc ctaatctctt 1021 tctttcaggg caataatgat acaatgtatc atgcctcttt gcaccattct aaagaataac 1081 agtgataatt tctgggttaa ggcaatagca atatttctgc atataaatat ttctgcatat 1141 aaattgtaac tgatgtaaga ggtttcatat tgctaatagc agctacaatc cagctaccat 1201 tctgctttta ttttatggtt gggataaggc tggattattc tgagtccaag ctaggccctt 1261 ttgctaatca tgttcatacc tcttatcttc ctcccacagc tcctgggcaa cgtgctggtc 1321 tgtgtgctgg cccatcactt tggcaaagaa ttgggattcg aacatcgatt gaattccccg 1381 gggatcctct agagtcgacc tgcagaagct atggcacccc tgaggccctt tttcatacta 1441 gccctggtgg catgggtttc tctggctgac caagagtcat gcaagggccg ctgcactcag 1501 ggtttcatgg ccagcaagaa gtgtcagtgt gacgagcttt gcacttacta tcagagctgc 1561 tgtgccgact acatggagca gtgcaagccc caagtaacgc ggggggacgt gttcactatg 1621 ccagaggatg attattggag ctatgactac gtggaggagc ccaagaacaa taccaacacc 1681 ggtgtgcaac ccgagaacac ctctccaccc ggtgacctaa atcctcggac ggacggcact 1741 ctaaagccga cagccttcct agatcctgag gaacagccaa gcaccccagc gcctaaagtg 1801 gagcaacagg aggagatcct aaggcccgac accactgatc aagggacccc tgagtttcca 1861 gaggaagaac tgtgcagtgg aaagcccttt gacgccttca cggatctcaa gaatgggtcc 1921 ctctttgcct tccgagggca gtactgctat gagctagatg agacggcagt gaggcctggg 1981 taccccaaac ttatccaaga tgtctggggc attgagggcc ccatcgatgc tgccttcact 2041 cgcatcaact gtcaggggaa gacctacttg ttcaagggta gtcagtactg gcgctttgag 2101 gatggggtcc tggaccctgg ttatccccga aacatctccg aaggcttcag tggcatacca 2161 gacaatgttg atgcagcgtt cgcccttcct gcccaccgtt acagtggccg ggaaagggtc 2221 tacttcttca aggggaagca gtactgggag tacgaatttc agcagcaacc cagccaggag 2281 gagtgcgaag gcagctctct gtcagccgtg tttgagcact ttgccttgct tcagcgggac 2341 agctgggaga acattttcga actcctcttc tggggcagat cctctgatgg agccagagaa 2401 ccccaattca tcagccggaa ctggcatggt gtgccaggga aagtggacgc tgctatggcc 2461 ggccgcatct acgtcactgg ctccttatcc cactctgccc aagccaaaaa acagaagtct 2521 aagcgtagaa gccgaaagcg ctatcgttca cgccgagggc gtggccacag acgcagccag 2581 agctcgaact cccgtcgttc atcacgttca atctggttct ctttgttctc cagcgaggag 2641 agtgggctag gaacctacaa caactatgat tatgatatgg actggcttgt acctgccacc 2701 tgcgagccca ttcagagcgt ctatttcttc tctggagaca aatactaccg agtcaacctt 2761 agaacccggc gagtggactc tgtgaatcct ccctacccac gctccattgc tcagtattgg 2821 ctgggctgcc cgacctctga gaaagcttac ccatacgatg ttccagatta cgcttaggat 2881 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac 2941 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt 3001 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa 3061 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc 3121 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga 3181 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag 3241 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc 3301 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct 3361 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag 3421 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 3481 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag 3541 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt 3601 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 3661 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc 3721 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 3781 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg 3841 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 3901 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct 3961 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga 4021 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca 4081 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac 4141 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga 4201 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac 4261 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 4321 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 4381 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 4441 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 4501 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 4561 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 4621 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 4681 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 4741 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 4801 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 4861 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 4921 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 4981 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 5041 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 5101 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 5161 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 5221 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 5281 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 5341 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 5401 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 5461 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 5521 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 5581 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 5641 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 5701 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 5761 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 5821 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 5881 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 5941 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 6001 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 6061 ggcggagcct atggaaaaac gccagcaacg //