LOCUS       Exported                6515 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses murine Thrombospondin-4 (THBS4) protein with HA-tag at 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CMV.SV40.THBS4-HA.SV40(polyA)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6515)
  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 6515)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6515
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   44..173
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        326..529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     480..500
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     526..550
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3607..3633
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3690..3709
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3821..3961
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3821..3950
                     /label=AAV2 ITR
     rep_origin      4036..4491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4123..4142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4333..4354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4508..4527)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4627..4649
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4687..4705)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4773..4877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4878..5738
                     /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(5096..5115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5909..6497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6398..6417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
      181 ctcgtgacct aggcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      241 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      301 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      361 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      421 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      481 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      541 ccgtcagatc gcctggagac gccatccgga ctctaaggta aatataaaat ttttaagtgt
      601 ataatgtgtt aaactactga ttctaattgt ttctctcttt tagattccaa cctttggaac
      661 tgaattccgc gggcccggga tcctctagag tcgacctgca gaagctgagt caccatgccg
      721 gccccacgcg cggccgcggc cgccttcctc ctgctgcacc tggtcctgca gccgtggcag
      781 cggacgagcg cccaggccac accccaggtc tttgatcttc taccgtcctc cagccagagg
      841 ctgaacccca gcgccctgca gccagtcctg acagatccca ccctgcacga agtctatctc
      901 atctccacct tcaagctgca gagtaaaagc tcagccacca tctttggcct ctactcctca
      961 agtgacaaca gcaaatactt tgaattcact gtgatgggac ggctgaacaa agccatcctc
     1021 cgctacctga agaatgatgg aaagatccat ctggtggtgt tcaacaactt gcagctggct
     1081 gatgggaggc gccacagggt tctcctgaga ctgagcaatt tgcagagagg agatggctct
     1141 gtagaactct acctggactg cgctcaggcg gactcggtgc gcaacctccc cagagccttt
     1201 tctggcctca cccagaaccc agagtccatt gaattgcgga cattccagag gaagccccag
     1261 gacttcttgg aagagctgaa gctggtggtg cggggctcgc tgttccaggt ggccagcctc
     1321 caagactgct tcctgcagca gagtgagcct ctggctgcca caagcacagg agactttaat
     1381 cgacagttct tgggtcaaat gacacaactg aaccagctcc ttggagaggt gaaagatctt
     1441 ctgaggcagc aggtcaagga aacatcgttt ttgcgaaaca ccattgctga atgccaagct
     1501 tgtggtcctc tcagctttca gtccccaact ccaaacacac tggtgcccat agctcctcca
     1561 gcacccccaa ctcgcccaac acgccactgt gactccagcc cttgtttccg aggtgtccga
     1621 tgtacggaca ccagggatgg cttccagtgt ggaccctgcc cagatggtta cacgggaaac
     1681 gggatcacct gctctgacgt ggatgagtgc aaataccatc cctgctatcc aggtgtgcgc
     1741 tgtgtgaatt tggctccggg tttcagatgt gacgcctgtc cagtaggttt cacagggccc
     1801 atggtgcagg gtgtcgggat caactttgct aaaacaaaca agcaggtgtg cactgatgtt
     1861 gatgagtgtc agaatggagc gtgtgttctg aattctatct gcattaacac tttgggatct
     1921 tatcgctgtg ggccttgcaa accagggtac actggtgatc agacgagggg atgcaagact
     1981 gaaaggagct gccggaaccc agagcagaac ccatgcagtg tgcatgcgca gtgcattgag
     2041 gagcggcaag gggatgtgac gtgtgtgtgt ggtgttggct gggctggtga cggctatgtc
     2101 tgtggaaagg atgtggacat tgacagttac cctgatgaag aactgccatg ctctgccagg
     2161 aactgcaaga aggataactg caagtatgtg ccaaactctg gccaggaaga tgcagacaga
     2221 gatggcatcg gagacgcgtg tgatgaggat gcagatggag atgggatctt gaatgaacag
     2281 gacaactgtg tcctgactca caatatagac caaaggaaca gtgataaaga tatctttggg
     2341 gatgcctgtg acaactgccg gatggtcctg aataatgacc agaaggatac tgacggggat
     2401 gggagaggag acgcctgtga tgacgacatg gatggagatg gaataaaaaa cattttggac
     2461 aactgtccca gagttcccaa ccgtgaccag caggatcggg atggtgatga tgtcggagat
     2521 gcttgtgaca gctgtcctga tgtcagtaac cctaaccagt cggatgtgga taacgatctg
     2581 gttggggact cctgtgacac caaccaagac agtgacgggg atggccacca ggacagcaca
     2641 gacaactgcc ccacagtcat taacagctcc cagctggaca ctgacaaaga tgggatcggc
     2701 gatgaatgtg atgacgatga cgataacgat ggcatcccag acctggtgcc tcctggccca
     2761 gacaactgca ggctcgtccc caacccagcc caggaagata gcaacaatga cggggtagga
     2821 gatatctgtg aagctgactt cgaccaggac caggtcatcg atcacattga tgtgtgccct
     2881 gagaatgcgg agatcaccct gactgacttc agggcctacc agacagtggt cctggaccct
     2941 gaaggggatg ctcagatcga ccccaactgg gtggtcctga accagggcat ggaaattgtg
     3001 cagaccatga acagtgaccc tggcctagct gttgggtaca cggcatttaa cggagttgac
     3061 tttgaaggga ccttccacgt aaacacccag acagatgatg actatgctgg ctttattttt
     3121 ggttaccaag acagctccag cttctatgta gtcatgtgga agcaaacgga gcagacatac
     3181 tggcaggcta caccgttccg tgcagtagca gaacctggta ttcagctcaa ggctgtgaag
     3241 tctaaaactg gcccagggga acatctccga aactccctgt ggcacacagg ggacaccagc
     3301 gaccaggtca ggctgctgtg gaaggactcc aggaatgtgg gctggaagga caaggtctcc
     3361 taccgctggt tcctgcagca taggccccag gtgggctaca tcagggtgcg cttttatgaa
     3421 ggctcggagt tggtggctga ctctggggtc accatagaca ccaccatgcg tggaggccga
     3481 cttggtgtgt tctgcttctc tcaagaaaac atcatctggt ccaacctcaa gtaccgctgc
     3541 aacgacacca taccagagga cttccaggag ttccagaccc agagtttcga ccgcttggat
     3601 aaagcttacc catacgatgt tccagattac gcttagctcg agagatctag gccgctaggc
     3661 ctcacctgcg atctcgatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3721 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3781 ttcaggggga ggtgtgggag gttttttaaa ctgcggccgc aggaacccct agtgatggag
     3841 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3901 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3961 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     4021 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4081 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     4141 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4201 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     4261 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4321 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     4381 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4441 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4501 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4561 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4621 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4681 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4741 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4801 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4861 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4921 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4981 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5041 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5101 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5161 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5221 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5281 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5341 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5401 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5461 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5521 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5581 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5641 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5701 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5761 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5821 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5881 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5941 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6001 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6061 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6121 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6181 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6241 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6301 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6361 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6421 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6481 ggcggagcct atggaaaaac gccagcaacg cggcc
//
