LOCUS       Exported                7147 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses murine Thrombospondin-1 (THBS1) protein with HA-tag at 
            C-terminus and KDEL (ER retention) sequence.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CMV.SV40.THBS1-HA-KDEL.SV40(polyA)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7147)
  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 7147)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7147
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   47..176
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..503
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     529..553
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4230..4256
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     4325..4344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4456..4596
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4456..4585
                     /label=AAV2 ITR
     rep_origin      4671..5126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4758..4777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4968..4989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5143..5162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5262..5284
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5322..5340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5408..5512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5513..6373
                     /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(5731..5750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6544..7132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7033..7052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
       61 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
      121 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
      181 ccgctcgtga cctaggcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
      541 gaaccgtcag atcgcctgga gacgccatcc ggactctaag gtaaatataa aatttttaag
      601 tgtataatgt gttaaactac tgattctaat tgtttctctc ttttagattc caacctttgg
      661 aactgaattc cgcgggcccg ggatcctcta gagtcgacct gcagaagctt atgaccatgg
      721 agctcctgcg gggactaggt gtcctgttcc tgttgcatat gtgtggaagc aaccgcattc
      781 cagagtctgg gggagataac ggtgtgtttg acatctttga actcattgga ggtgcacgaa
      841 ggggccccgg tcgccgactg gtgaagggcc aagatctatc cagccccgcc ttccgcattg
      901 agaatgccaa cctgatcccc gctgtgccgg atgacaagtt ccaagaccta ctggacgctg
      961 tgtgggccga caaaggcttc atcttcctgg cttccttgag gcagatgaag aagacccggg
     1021 gcacactcct ggctgtggaa cggaaagaca acactggcca gatcttcagt gtggtctcca
     1081 acggcaaagc tggcaccctg gacctgagcc tgagcctgcc agggaagcaa caagtggtgt
     1141 cagtggagga agctctcctg gccactggcc agtggaagag catcacgctg tttgttcaag
     1201 aggaccgggc tcaactctac atagactgtg ataagatgga gagcgcggag ctggatgtac
     1261 ccatccagag catcttcacc agggatctgg ccagcgttgc caggctccga gttgcaaagg
     1321 gagatgtcaa tgacaatttt cagggggtgc tgcagaatgt gaggtttgtc tttggaacca
     1381 ccccagaaga cattctcagg aacaaaggct gctccagctc agctaccaac gtccttctta
     1441 cccttgacaa caacgtggtg aacggttcca gccctgctat ccgcaccaac tacatcggcc
     1501 acaaaacaaa ggacctccaa gctatctgtg gcctctcctg tgatgaacta tccagcatgg
     1561 tcctggaact gaagggcctg cgcaccatcg tgaccactct gcaggacagc atccgaaaag
     1621 tgacggaaga gaacagagag ctggtcagtg agctgaagcg gcctcccctc tgctttcaca
     1681 atggagtcca gtacaagaac aacgaggagt ggactgtaga cagttgcaca gagtgtcact
     1741 gccagaactc ggttaccatc tgcaaaaagg tgtcctgtcc catcatgccc tgctccaacg
     1801 ccacagttcc tgatggtgaa tgctgcccac ggtgctggcc cagcgactct gctgacgatg
     1861 gctggtctcc ctggtctgag tggacctcct gctctgccac atgtggcaat ggaattcagc
     1921 aacgtggtcg ttcctgtgac agcctcaaca acagatgcga gggctcttcg gtacagacaa
     1981 ggacctgcca cattcaggag tgtgacaaaa gatttaaaca ggatggtggc tggagtcact
     2041 ggtctccatg gtcgtcctgt tctgtgacct gtggtgacgg tgtgatcaca aggatccggc
     2101 tctgcaactc ccccagcccc cagatgaacg ggaagccctg tgaaggtgaa gcccgggaga
     2161 ccaaagcctg caagaaagac gcctgcccaa ttaatggagg ctggggtccc tggtcaccat
     2221 gggacatctg ctctgtcacc tgtggaggag gagtgcagag acgcagccga ctctgtaaca
     2281 accccacacc ccagtttgga ggcaaagact gtgttggcga tgtgacagaa aatcaagttt
     2341 gcaacaagca ggactgccca attgatggat gcctgtccaa tccctgcttt gctggtgcca
     2401 agtgtactag ctaccctgat ggtagctgga aatgtggtgc gtgtcctcct ggctacagtg
     2461 gaaatggcat ccagtgcaaa gacgtcgatg agtgcaaaga agtgcctgat gcttgcttca
     2521 atcacaacgg agaacatcgg tgcaagaaca cagatcctgg ctacaactgc ctgccctgcc
     2581 caccacgatt cactggctca cagcccttcg gccgaggtgt cgaacatgcc atggccaaca
     2641 aacaggtgtg caaaccgcga aacccctgca cggacgggac gcatgactgc aacaagaacg
     2701 ctaagtgcaa ctacctgggt cactacagtg accccatgta ccgctgtgag tgcaagcccg
     2761 gctatgcagg caatggcatc atctgcggag aggacacaga cctggacggc tggcctaatg
     2821 aaaacctggt gtgtgtggcc aacgcaacct accactgcaa aaaggacaac tgccccaacc
     2881 ttcccaactc ggggcaggaa gactatgaca aggacgggat tggcgatgcc tgcgatgatg
     2941 acgatgacaa cgacaagatc cccgatgaca gggacaactg tccattccat tacaacccag
     3001 cccagtatga ctatgacaga gatgatgtgg gagaccgctg tgacaactgc ccctacaacc
     3061 acaaccctga ccaagcagac acagacaaaa acggggaggg cgatgcctgt gctgtggaca
     3121 tcgatggaga tggaatcctc aatgaacgag acaactgcca gtacgtttac aacgtggacc
     3181 agagggacac ggacatggat ggggttggag atcagtgtga caactgcccc ctggaacaca
     3241 atccagacca gctggactct gactcagacc tcatagggga cacttgtgac aacaatcagg
     3301 acatcgatga ggatggccat cagaacaatc tggacaactg tccctatgtg cctaatgcca
     3361 accaggccga ccatgataaa gatggcaaag gagatgcctg tgaccatgac gatgacaatg
     3421 acggcatccc tgatgacaga gacaactgca ggctggtgcc caatcctgac cagaaggact
     3481 ctgatggtga tggccgaggt gacgcctgca aagacgactt tgaccatgac aatgtgccag
     3541 atattgatga catctgtcct gagaattttg acatcagtga aaccgatttc cgacaattcc
     3601 agatgattcc tctagatccc aaaggaacct cccaaaatga ccctaactgg gttgtccgcc
     3661 atcagggcaa agaacttgtc cagactgtaa actgtgaccc tggacttgct gtaggttatg
     3721 atgagtttaa tgctgtggac ttcagcggta ccttcttcat caacaccgag agagatgatg
     3781 actacgctgg ctttgttttc ggctaccagt ccagcagccg cttctacgtt gtgatgtgga
     3841 aacaagtcac ccagtcctac tgggacacca accccacaag ggctcaggga tactcaggcc
     3901 tgtctgtaaa ggttgtgaac tccaccaccg gccctggcga gcacctgcgg aatgcactgt
     3961 ggcacacagg aaacacccct ggccaggtgc gcaccctgtg gcatgaccct cgccacatcg
     4021 gctggaaaga tttcactgca tacagatggc gtctcagcca caggccaaag accggttata
     4081 tcagagtggt gatgtatgaa ggaaagaaaa tcatggctga ctcgggaccc atctatgaca
     4141 aaacctacgc cggcggtaga ctaggcctgt tcgtcttctc tcaggaaatg gtgttcttct
     4201 cagacatgaa atacgagtgt cgagattcct acccatacga tgttccagat tacgctaaag
     4261 acgaactgta actcgagaga tctaggccgc taggcctcac ctgcgatctc gatgctttat
     4321 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     4381 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     4441 ttaaactgcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4501 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4561 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4621 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4681 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4741 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4801 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4861 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4921 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4981 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     5041 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5101 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5161 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5221 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5281 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5341 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7141 caacgcg
//
