LOCUS       Exported                7419 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Truncated Mouse gamma-synuclein (mSncg-0.27kb) promoter-mediates HA 
            and Cas9 expression in retinal ganglion cells.
ACCESSION   .
VERSION     .
KEYWORDS    px551-mSncg-0.27kb-HA-SpCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  AUTHORS   Wang Q, Zhuang PS, Huang H, Li L, Liu L, Webber HC, Dalal R, Siew L,
            Fligor CM, Chang KC, Nahmou M, Kreymerman A, Sun Y, Meyer JS, 
            Goldberg JL, Hu Y
  TITLE     Mouse gamma-Synuclein Promoter-Mediated Gene Expression and Editing 
            in Mammalian Retinal Ganglion Cells.
  JOURNAL   J Neurosci. 2020 Apr 9. pii: JNEUROSCI.0102-20.2020. doi: 
            10.1523/JNEUROSCI.0102-20.2020.
  PUBMED    32300046
REFERENCE   2  (bases 1 to 7419)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7419
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             423..449
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             453..473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             483..4583
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4587..4607
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4626..4674
                     /note="synthetic polyadenylation signal"
     repeat_region   4682..4822
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4682..4811
                     /label=AAV2 ITR
     rep_origin      4897..5352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4984..5003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5194..5215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5369..5388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5488..5510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5548..5566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5634..5738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5739..6599
                     /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(5957..5976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6770..7358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7259..7278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gaaagagtca gcagggcaga atagagccag taggggcccg
      181 ggccctgctc gctggtatcc ccgtgaggca tgccttctct ctggcccgcc ctccctgccc
      241 ccaccctggc ccgggctggc tgggctccag ccagcagcca cagcatcaat atttcatctg
      301 cgtcaataag aggcagtagc agcagagaca gcggctgcgg cagcactcca gtccatagct
      361 tgcagcagcc aggttccatc cttgcaaaca ccatggacgt cttcaagacc ggtgccacca
      421 tgtacccata cgatgttcca gattacgctt cgccgaagaa aaagcgcaag gtcgaagcgt
      481 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
      541 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
      601 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
      661 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
      721 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
      781 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
      841 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
      901 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
      961 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     1021 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     1081 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     1141 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     1201 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     1261 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1321 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1381 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     1441 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     1501 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     1561 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     1621 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     1681 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     1741 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     1801 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     1861 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     1921 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     1981 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     2041 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     2101 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     2161 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     2221 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     2281 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     2341 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     2401 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     2461 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     2521 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     2581 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     2641 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     2701 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     2761 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     2821 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     2881 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     2941 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     3001 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     3061 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     3121 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     3181 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     3241 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     3301 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     3361 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     3421 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     3481 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     3541 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     3601 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     3661 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     3721 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     3781 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     3841 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     3901 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     3961 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     4021 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     4081 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     4141 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     4201 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     4261 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     4321 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     4381 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     4441 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     4501 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     4561 acctgtctca gctgggaggc gacagcccca agaagaagag aaaggtggag gccagctaag
     4621 aattcaataa aagatcttta ttttcattag atctgtgtgt tggttttttg tgtgcggccg
     4681 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4741 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4801 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4861 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4921 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4981 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5041 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5101 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5161 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5221 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     5281 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5341 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5401 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5461 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5521 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5581 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5641 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5701 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5761 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5821 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5881 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5941 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6001 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6061 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6121 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6181 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6241 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6301 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6361 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6421 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
     6481 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6541 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6601 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6661 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6721 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6781 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6841 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6901 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6961 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7021 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7081 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7141 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7201 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7261 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7321 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7381 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
