LOCUS       Exported                7135 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses murine Thrombospondin-1 (THBS1) protein with HA-tag at 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.CMV.SV40.THBS1-HA.SV40(polyA)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7135)
  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 7135)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7135
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     3..27
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3704..3730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     3787..3806
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3918..4058
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3918..4047
                     /label=AAV2 ITR
     rep_origin      4133..4588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4220..4239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4430..4451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4605..4624)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4724..4746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4784..4802)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4870..4974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4975..5835
                     /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(5193..5212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6006..6594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6495..6514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6656..6785
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        6938..6
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7092..7112
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccggactc taaggtaaat
       61 ataaaatttt taagtgtata atgtgttaaa ctactgattc taattgtttc tctcttttag
      121 attccaacct ttggaactga attccgcggg cccgggatcc tctagagtcg acctgcagaa
      181 gcttatgacc atggagctcc tgcggggact aggtgtcctg ttcctgttgc atatgtgtgg
      241 aagcaaccgc attccagagt ctgggggaga taacggtgtg tttgacatct ttgaactcat
      301 tggaggtgca cgaaggggcc ccggtcgccg actggtgaag ggccaagatc tatccagccc
      361 cgccttccgc attgagaatg ccaacctgat ccccgctgtg ccggatgaca agttccaaga
      421 cctactggac gctgtgtggg ccgacaaagg cttcatcttc ctggcttcct tgaggcagat
      481 gaagaagacc cggggcacac tcctggctgt ggaacggaaa gacaacactg gccagatctt
      541 cagtgtggtc tccaacggca aagctggcac cctggacctg agcctgagcc tgccagggaa
      601 gcaacaagtg gtgtcagtgg aggaagctct cctggccact ggccagtgga agagcatcac
      661 gctgtttgtt caagaggacc gggctcaact ctacatagac tgtgataaga tggagagcgc
      721 ggagctggat gtacccatcc agagcatctt caccagggat ctggccagcg ttgccaggct
      781 ccgagttgca aagggagatg tcaatgacaa ttttcagggg gtgctgcaga atgtgaggtt
      841 tgtctttgga accaccccag aagacattct caggaacaaa ggctgctcca gctcagctac
      901 caacgtcctt cttacccttg acaacaacgt ggtgaacggt tccagccctg ctatccgcac
      961 caactacatc ggccacaaaa caaaggacct ccaagctatc tgtggcctct cctgtgatga
     1021 actatccagc atggtcctgg aactgaaggg cctgcgcacc atcgtgacca ctctgcagga
     1081 cagcatccga aaagtgacgg aagagaacag agagctggtc agtgagctga agcggcctcc
     1141 cctctgcttt cacaatggag tccagtacaa gaacaacgag gagtggactg tagacagttg
     1201 cacagagtgt cactgccaga actcggttac catctgcaaa aaggtgtcct gtcccatcat
     1261 gccctgctcc aacgccacag ttcctgatgg tgaatgctgc ccacggtgct ggcccagcga
     1321 ctctgctgac gatggctggt ctccctggtc tgagtggacc tcctgctctg ccacatgtgg
     1381 caatggaatt cagcaacgtg gtcgttcctg tgacagcctc aacaacagat gcgagggctc
     1441 ttcggtacag acaaggacct gccacattca ggagtgtgac aaaagattta aacaggatgg
     1501 tggctggagt cactggtctc catggtcgtc ctgttctgtg acctgtggtg acggtgtgat
     1561 cacaaggatc cggctctgca actcccccag cccccagatg aacgggaagc cctgtgaagg
     1621 tgaagcccgg gagaccaaag cctgcaagaa agacgcctgc ccaattaatg gaggctgggg
     1681 tccctggtca ccatgggaca tctgctctgt cacctgtgga ggaggagtgc agagacgcag
     1741 ccgactctgt aacaacccca caccccagtt tggaggcaaa gactgtgttg gcgatgtgac
     1801 agaaaatcaa gtttgcaaca agcaggactg cccaattgat ggatgcctgt ccaatccctg
     1861 ctttgctggt gccaagtgta ctagctaccc tgatggtagc tggaaatgtg gtgcgtgtcc
     1921 tcctggctac agtggaaatg gcatccagtg caaagacgtc gatgagtgca aagaagtgcc
     1981 tgatgcttgc ttcaatcaca acggagaaca tcggtgcaag aacacagatc ctggctacaa
     2041 ctgcctgccc tgcccaccac gattcactgg ctcacagccc ttcggccgag gtgtcgaaca
     2101 tgccatggcc aacaaacagg tgtgcaaacc gcgaaacccc tgcacggacg ggacgcatga
     2161 ctgcaacaag aacgctaagt gcaactacct gggtcactac agtgacccca tgtaccgctg
     2221 tgagtgcaag cccggctatg caggcaatgg catcatctgc ggagaggaca cagacctgga
     2281 cggctggcct aatgaaaacc tggtgtgtgt ggccaacgca acctaccact gcaaaaagga
     2341 caactgcccc aaccttccca actcggggca ggaagactat gacaaggacg ggattggcga
     2401 tgcctgcgat gatgacgatg acaacgacaa gatccccgat gacagggaca actgtccatt
     2461 ccattacaac ccagcccagt atgactatga cagagatgat gtgggagacc gctgtgacaa
     2521 ctgcccctac aaccacaacc ctgaccaagc agacacagac aaaaacgggg agggcgatgc
     2581 ctgtgctgtg gacatcgatg gagatggaat cctcaatgaa cgagacaact gccagtacgt
     2641 ttacaacgtg gaccagaggg acacggacat ggatggggtt ggagatcagt gtgacaactg
     2701 ccccctggaa cacaatccag accagctgga ctctgactca gacctcatag gggacacttg
     2761 tgacaacaat caggacatcg atgaggatgg ccatcagaac aatctggaca actgtcccta
     2821 tgtgcctaat gccaaccagg ccgaccatga taaagatggc aaaggagatg cctgtgacca
     2881 tgacgatgac aatgacggca tccctgatga cagagacaac tgcaggctgg tgcccaatcc
     2941 tgaccagaag gactctgatg gtgatggccg aggtgacgcc tgcaaagacg actttgacca
     3001 tgacaatgtg ccagatattg atgacatctg tcctgagaat tttgacatca gtgaaaccga
     3061 tttccgacaa ttccagatga ttcctctaga tcccaaagga acctcccaaa atgaccctaa
     3121 ctgggttgtc cgccatcagg gcaaagaact tgtccagact gtaaactgtg accctggact
     3181 tgctgtaggt tatgatgagt ttaatgctgt ggacttcagc ggtaccttct tcatcaacac
     3241 cgagagagat gatgactacg ctggctttgt tttcggctac cagtccagca gccgcttcta
     3301 cgttgtgatg tggaaacaag tcacccagtc ctactgggac accaacccca caagggctca
     3361 gggatactca ggcctgtctg taaaggttgt gaactccacc accggccctg gcgagcacct
     3421 gcggaatgca ctgtggcaca caggaaacac ccctggccag gtgcgcaccc tgtggcatga
     3481 ccctcgccac atcggctgga aagatttcac tgcatacaga tggcgtctca gccacaggcc
     3541 aaagaccggt tatatcagag tggtgatgta tgaaggaaag aaaatcatgg ctgactcggg
     3601 acccatctat gacaaaacct acgccggcgg tagactaggc ctgttcgtct tctctcagga
     3661 aatggtgttc ttctcagaca tgaaatacga gtgtcgagat tcctacccat acgatgttcc
     3721 agattacgct taactcgaga gatctaggcc gctaggcctc acctgcgatc tcgatgcttt
     3781 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3841 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3901 ttttaaactg cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3961 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4021 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4081 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4141 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4201 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4261 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4321 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4381 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4441 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4501 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4561 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4621 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4681 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4741 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4801 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4861 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5101 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5161 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5221 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5281 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5341 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5401 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5461 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5521 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5581 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5641 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5701 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5761 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5821 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5881 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5941 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6061 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     6661 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     6721 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     6781 ttcctgcggc cgctcgtgac ctaggcatat gccaagtacg ccccctattg acgtcaatga
     6841 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     6901 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     6961 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7021 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     7081 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagc
//
