LOCUS       Exported                6771 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  LITE1.0 CRY2PHR fused to VP64 transcriptional activator domain. 
            Binds to CIB1 upon blue light stimulation. Synapsin promoter for 
            neuronal expression. See LITE2.0 for optimized LITE activators..
ACCESSION   .
VERSION     .
KEYWORDS    LITE1.0_pAAV_hSyn_CRY2PHR-NLS-VP64_2A_GFP_WPRE_bGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6771)
  AUTHORS   Konermann S, Brigham MD, Trevino A, Hsu PD, Heidenreich M, Le Cong, 
            Platt RJ, Scott DA, Church GM, Zhang F
  TITLE     Optical control of mammalian endogenous transcription and epigenetic
            states.
  JOURNAL   Nature. 2013 Jul 23. doi: 10.1038/nature12466.
  PUBMED    23877069
REFERENCE   2  (bases 1 to 6771)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6771
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2151..2171
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2196..2345
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             2367..2420
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2421..3137
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             2421..3134
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2463..2484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2724..2743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3071..3092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3162..3750
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3215..3235)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3633..3644)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3792..3809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3798..4005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4034..4174
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4034..4163
                     /label=AAV2 ITR
     rep_origin      4249..4704
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4336..4355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4546..4567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4721..4740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4840..4862
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4900..4918)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4986..5090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5091..5951
                     /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(5309..5328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6122..6710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6611..6630
                     /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 gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg aagatggaca
      661 aaaagacgat tgtgtggttt agacgtgacc ttcgcattga ggacaaccct gcgttggcag
      721 cggcagcgca tgaaggttcg gtattccctg tctttatctg gtgtcccgaa gaggaggggc
      781 aattctatcc cggtcgcgcg tcccgatggt ggatgaaaca gtcgctcgca catcttagcc
      841 agtcgttgaa ggcgctcggc tctgatctga cgctcatcaa aacacacaat acaatttctg
      901 ccattctgga ttgcattagg gtgactggtg ccaccaaagt tgtgttcaat cacctttatg
      961 atcccgtaag cctggtcagg gatcatacag taaaagagaa acttgtagag aggggaatca
     1021 gcgtccaatc gtacaatggg gacttgttgt atgaaccgtg ggaaatctat tgtgagaaag
     1081 gcaaaccttt cacttccttc aattcgtact ggaagaaatg cctcgacatg tccatcgagt
     1141 cagtcatgct gccgccacct tggagactta tgcccattac tgctgcggca gaggcgatct
     1201 gggcttgcag catcgaagaa cttggtctgg aaaacgaagc ggaaaaaccc agtaacgcgc
     1261 tcctgacgcg agcgtggtcg ccaggctggt cgaatgctga taagctcctt aatgagttta
     1321 tcgagaaaca gttgatcgac tatgcgaaga atagcaagaa ggtggtggga aattccactt
     1381 ccctcctgtc gccgtatctc cactttgggg agattagcgt gaggcatgtg ttccaatgtg
     1441 cgcggatgaa gcaaatcatt tgggctcgcg acaaaaactc tgaaggagag gagtctgccg
     1501 atttgttttt gagaggaatc ggccttaggg agtattcacg atacatctgc ttcaactttc
     1561 cgtttacaca tgaacaatcc ctcctcagcc acctccgctt ctttccgtgg gacgcggacg
     1621 tggacaaatt caaggcctgg cggcaggggc gcacgggata cccgttggtg gatgcgggta
     1681 tgagagagct gtgggcaacg ggttggatgc acaaccgtat tcgagtcatc gttagtagct
     1741 ttgccgttaa gttcctcttg ttgccctgga aatgggggat gaaatacttt tgggacacgc
     1801 ttctggatgc ggacttggag tgcgatattc ttgggtggca gtacatttcg ggttccatcc
     1861 ctgacgggca tgaacttgac cgattggata accctgcact gcaaggagca aagtacgacc
     1921 cggagggaga atacatcagg cagtggctcc cagagctggc ccgtctgccc accgagtgga
     1981 ttcaccaccc gtgggatgct cccctgaccg tccttaaagc ctcaggggtc gaattgggga
     2041 cgaattacgc gaaacccatc gtcgatattg acaccgcacg agaattgctg gcgaaagcca
     2101 tctcaagaac acgcgaagcg cagattatga tcggagcagc gccggctagc cccaagaaga
     2161 agagaaaggt ggaggccagc ggttccggac gggctgacgc attggacgat tttgatctgg
     2221 atatgctggg aagtgacgcc ctcgatgatt ttgaccttga catgcttggt tcggatgccc
     2281 ttgatgactt tgacctcgac atgctcggca gtgacgccct tgatgatttc gacctggaca
     2341 tgctgattaa ctctagaggc agtggagagg gcagaggaag tctgctaaca tgcggtgacg
     2401 tcgaggagaa tcctggccca gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     2461 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2521 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2581 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2641 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2701 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2761 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2821 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2881 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2941 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3001 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3061 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3121 acgagctgta caagtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca
     3181 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3241 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3301 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3361 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3421 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3481 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3541 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     3601 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3661 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3721 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagctagagc
     3781 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggaggtaa ccacgtgcgg
     4021 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4081 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4141 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     4201 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     4261 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4321 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4381 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4441 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4501 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4561 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     4621 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4681 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4741 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4801 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4861 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4921 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5641 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5701 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5761 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     5821 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5881 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5941 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6001 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6061 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6241 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
