LOCUS       Exported                7015 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses murine Thrombospondin-1 (THBS1) protein with deletion of 
            Coiled Coil domain and HA-tag at C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CMV.SV40.THBS1-deltaCC-HA.SV40(polyA)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7015)
  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 7015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7015
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        59..262
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     213..233
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     259..283
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3840..3866
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3923..3942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4054..4194
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4054..4183
                     /label=AAV2 ITR
     rep_origin      4269..4724
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4356..4375)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4566..4587
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4741..4760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4860..4882
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4920..4938)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5006..5110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5111..5971
                     /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(5329..5348)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6142..6730
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6631..6650
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6792..6921
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
       61 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      121 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      181 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      241 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
      301 ggactctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
      361 tgtttctctc ttttagattc caacctttgg aactgaattc cgcgggcccg ggatcctcta
      421 gagtcgacct gcagaagctt atgaccatgg agctcctgcg gggactaggt gtcctgttcc
      481 tgttgcatat gtgtggaagc aaccgcattc cagagtctgg gggagataac ggtgtgtttg
      541 acatctttga actcattgga ggtgcacgaa ggggccccgg tcgccgactg gtgaagggcc
      601 aagatctatc cagccccgcc ttccgcattg agaatgccaa cctgatcccc gctgtgccgg
      661 atgacaagtt ccaagaccta ctggacgctg tgtgggccga caaaggcttc atcttcctgg
      721 cttccttgag gcagatgaag aagacccggg gcacactcct ggctgtggaa cggaaagaca
      781 acactggcca gatcttcagt gtggtctcca acggcaaagc tggcaccctg gacctgagcc
      841 tgagcctgcc agggaagcaa caagtggtgt cagtggagga agctctcctg gccactggcc
      901 agtggaagag catcacgctg tttgttcaag aggaccgggc tcaactctac atagactgtg
      961 ataagatgga gagcgcggag ctggatgtac ccatccagag catcttcacc agggatctgg
     1021 ccagcgttgc caggctccga gttgcaaagg gagatgtcaa tgacaatttt cagggggtgc
     1081 tgcagaatgt gaggtttgtc tttggaacca ccccagaaga cattctcagg aacaaaggct
     1141 gctccagctc agctaccaac gtccttctta cccttgacaa caacgtggtg aacggttcca
     1201 gccctgctat ccgcaccaac tacatcggcc acaaaacaaa ggacctccaa gctatctgtg
     1261 gcctctcctg tcctcccctc tgctttcaca atggagtcca gtacaagaac aacgaggagt
     1321 ggactgtaga cagttgcaca gagtgtcact gccagaactc ggttaccatc tgcaaaaagg
     1381 tgtcctgtcc catcatgccc tgctccaacg ccacagttcc tgatggtgaa tgctgcccac
     1441 ggtgctggcc cagcgactct gctgacgatg gctggtctcc ctggtctgag tggacctcct
     1501 gctctgccac atgtggcaat ggaattcagc aacgtggtcg ttcctgtgac agcctcaaca
     1561 acagatgcga gggctcttcg gtacagacaa ggacctgcca cattcaggag tgtgacaaaa
     1621 gatttaaaca ggatggtggc tggagtcact ggtctccatg gtcgtcctgt tctgtgacct
     1681 gtggtgacgg tgtgatcaca aggatccggc tctgcaactc ccccagcccc cagatgaacg
     1741 ggaagccctg tgaaggtgaa gcccgggaga ccaaagcctg caagaaagac gcctgcccaa
     1801 ttaatggagg ctggggtccc tggtcaccat gggacatctg ctctgtcacc tgtggaggag
     1861 gagtgcagag acgcagccga ctctgtaaca accccacacc ccagtttgga ggcaaagact
     1921 gtgttggcga tgtgacagaa aatcaagttt gcaacaagca ggactgccca attgatggat
     1981 gcctgtccaa tccctgcttt gctggtgcca agtgtactag ctaccctgat ggtagctgga
     2041 aatgtggtgc gtgtcctcct ggctacagtg gaaatggcat ccagtgcaaa gacgtcgatg
     2101 agtgcaaaga agtgcctgat gcttgcttca atcacaacgg agaacatcgg tgcaagaaca
     2161 cagatcctgg ctacaactgc ctgccctgcc caccacgatt cactggctca cagcccttcg
     2221 gccgaggtgt cgaacatgcc atggccaaca aacaggtgtg caaaccgcga aacccctgca
     2281 cggacgggac gcatgactgc aacaagaacg ctaagtgcaa ctacctgggt cactacagtg
     2341 accccatgta ccgctgtgag tgcaagcccg gctatgcagg caatggcatc atctgcggag
     2401 aggacacaga cctggacggc tggcctaatg aaaacctggt gtgtgtggcc aacgcaacct
     2461 accactgcaa aaaggacaac tgccccaacc ttcccaactc ggggcaggaa gactatgaca
     2521 aggacgggat tggcgatgcc tgcgatgatg acgatgacaa cgacaagatc cccgatgaca
     2581 gggacaactg tccattccat tacaacccag cccagtatga ctatgacaga gatgatgtgg
     2641 gagaccgctg tgacaactgc ccctacaacc acaaccctga ccaagcagac acagacaaaa
     2701 acggggaggg cgatgcctgt gctgtggaca tcgatggaga tggaatcctc aatgaacgag
     2761 acaactgcca gtacgtttac aacgtggacc agagggacac ggacatggat ggggttggag
     2821 atcagtgtga caactgcccc ctggaacaca atccagacca gctggactct gactcagacc
     2881 tcatagggga cacttgtgac aacaatcagg acatcgatga ggatggccat cagaacaatc
     2941 tggacaactg tccctatgtg cctaatgcca accaggccga ccatgataaa gatggcaaag
     3001 gagatgcctg tgaccatgac gatgacaatg acggcatccc tgatgacaga gacaactgca
     3061 ggctggtgcc caatcctgac cagaaggact ctgatggtga tggccgaggt gacgcctgca
     3121 aagacgactt tgaccatgac aatgtgccag atattgatga catctgtcct gagaattttg
     3181 acatcagtga aaccgatttc cgacaattcc agatgattcc tctagatccc aaaggaacct
     3241 cccaaaatga ccctaactgg gttgtccgcc atcagggcaa agaacttgtc cagactgtaa
     3301 actgtgaccc tggacttgct gtaggttatg atgagtttaa tgctgtggac ttcagcggta
     3361 ccttcttcat caacaccgag agagatgatg actacgctgg ctttgttttc ggctaccagt
     3421 ccagcagccg cttctacgtt gtgatgtgga aacaagtcac ccagtcctac tgggacacca
     3481 accccacaag ggctcaggga tactcaggcc tgtctgtaaa ggttgtgaac tccaccaccg
     3541 gccctggcga gcacctgcgg aatgcactgt ggcacacagg aaacacccct ggccaggtgc
     3601 gcaccctgtg gcatgaccct cgccacatcg gctggaaaga tttcactgca tacagatggc
     3661 gtctcagcca caggccaaag accggttata tcagagtggt gatgtatgaa ggaaagaaaa
     3721 tcatggctga ctcgggaccc atctatgaca aaacctacgc cggcggtaga ctaggcctgt
     3781 tcgtcttctc tcaggaaatg gtgttcttct cagacatgaa atacgagtgt cgagattcct
     3841 acccatacga tgttccagat tacgcttaac tcgagagatc taggccgcta ggcctcacct
     3901 gcgatctcga tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3961 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4021 ggaggtgtgg gaggtttttt aaactgcggc cgcaggaacc cctagtgatg gagttggcca
     4081 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4141 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4201 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4261 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4321 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     4381 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4441 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4501 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4561 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4621 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4681 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4741 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4801 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4861 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4921 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4981 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5041 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5101 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5161 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5221 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5281 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5341 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5401 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5461 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5521 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5581 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5641 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5701 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5761 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5821 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5881 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5941 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6001 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6061 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6121 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6181 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6241 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6301 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6361 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6421 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6481 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6541 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6601 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6661 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6721 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6781 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
     6841 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     6901 actccatcac taggggttcc tgcggccgct cgtgacctag gcatatgcca agtacgcccc
     6961 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgac
//
