LOCUS       Exported                5339 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV genome that expresses nuclear localized mTurquoise2 from the 
            hSyn1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    hSyn1-2xNLS-mTurquoise2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5339)
  AUTHORS   Challis RC, Ravindra Kumar S, Chan KY, Challis C, Beadle K, Jang MJ,
            Kim HM, Rajendran PS, Tompkins JD, Shivkumar K, Deverman BE, 
            Gradinaru V
  TITLE     Systemic AAV vectors for widespread and targeted gene delivery in 
            rodents.
  JOURNAL   Nat Protoc. 2019 Feb;14(2):379-414. doi: 10.1038/s41596-018-0097-3.
  PUBMED    30626963
REFERENCE   2  (bases 1 to 5339)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5339
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      647..656
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             662..682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      689..698
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             695..1411
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             695..1411
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(740..761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1001..1020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1348..1369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1418..1438
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    1466..2054
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1519..1539)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1937..1948)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2086..2562
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2285..2304)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2602..2742
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2602..2731
                     /label=AAV2 ITR
     rep_origin      2817..3272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2904..2923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3114..3135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3289..3308)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3408..3430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3468..3486)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3554..3658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3659..4519
                     /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(3877..3896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4690..5278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5179..5198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccgcca ccatgggaag
      661 cccaaagaag aagcgtaagg taggatccgc caccatggtg agcaagggcg aggagctgtt
      721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      781 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgt cctggggcgt
      901 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1021 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acttcagcga
     1141 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1201 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1261 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     1321 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1381 gatcactctc ggcatggacg agctgtacaa gggaagcccc aagaaaaagc ggaaggtgtg
     1441 agaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     1501 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     1561 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     1621 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     1681 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     1741 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     1801 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     1861 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     1921 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     1981 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     2041 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     2101 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     2161 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     2221 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     2281 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     2341 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     2401 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2461 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2521 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     2581 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2641 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2701 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     2761 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     2821 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2881 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2941 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3001 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3061 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3121 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     3181 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3241 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3301 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3361 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3421 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3481 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3541 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3601 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3661 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3721 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3781 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3841 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3901 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3961 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4021 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4081 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4141 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4201 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4261 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4321 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4381 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4441 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4501 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4561 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4621 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4681 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4741 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4801 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4861 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4921 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4981 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5041 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5101 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5161 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5221 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5281 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
