LOCUS       Exported                6594 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  epiLITE / LITE1.0 CRY2PHR fused to SID4X transcriptional repressor 
            effector domain. Binds to CIB1 upon blue light stimulation. Synapsin
            promoter for neuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    LITE1.0_pAAV_hSyn_CRY2PHR-NLS-SID4X_2A_phiLOV2.1_WPRE_bGHpA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6594)
  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 6594)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6594
                     /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             2154..2174
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2574..2627
                     /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"
     misc_feature    2985..3573
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3038..3058)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3456..3467)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3615..3632)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3621..3828
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3857..3997
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3857..3986
                     /label=AAV2 ITR
     rep_origin      4072..4527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4159..4178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4369..4390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4544..4563)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4663..4685
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4723..4741)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4809..4913
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4914..5774
                     /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(5132..5151)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5945..6533
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6434..6453
                     /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 agccccaaga
     2161 agaagagaaa ggtggaggcc agcatgaaca tccagatgct gctggaggcc gctgactacc
     2221 tggaacggag agagcgcgaa gccgagcacg gatatgcttc aatgctgccc ggaagcggca
     2281 tgaatattca gatgctgctg gaggctgctg attacctgga aaggcgcgaa cgggaggccg
     2341 aacatggcta tgcttccatg ctgcctgggt ctggaatgaa tatccaaatg ctgctggagg
     2401 cagccgatta cctggaacgg agagaaagag aagccgagca cggatacgcc agcatgctgc
     2461 caggcagcgg gatgaacata caaatgctgc tggaggctgc cgattacctg gagaggcgcg
     2521 agagagaagc tgaacatggc tatgcctcta tgctgccccc atggggcagt ggagagggca
     2581 gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatc gagaagtcct
     2641 tcgtaataac agatccaaga ctgcctgatt accccatcat ttttgccagc gacggttttc
     2701 tggagctcac cgaatattcc agggaggaaa tcatggggag aaacgccagg ttcctgcaag
     2761 ggcctgagac agaccaggcc accgttcaga agattcgcga tgctatccgc gatcagaggg
     2821 aaaccactgt gcagttaata aactacacta agtcaggcaa aaagttctgg aatctgcttc
     2881 atctccaacc tgtccgggat cgaaagggag ggctgcaata cttcatcggt gtgcagctgg
     2941 tgggaagtga tcatgtgtaa gaattcgata tcaagcttat cgataatcaa cctctggatt
     3001 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3061 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3121 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3181 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3241 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     3301 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     3361 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     3421 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3481 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3541 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagctag
     3601 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggagg taaccacgtg
     3841 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3901 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3961 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc
     4021 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct
     4081 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4141 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4201 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4261 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4321 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4381 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4441 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4501 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg
     4561 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4621 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4681 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4741 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4801 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4861 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4921 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4981 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5041 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5101 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5161 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5221 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5281 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5341 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5401 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5461 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5521 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5581 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta
     5641 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5701 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5761 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5821 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5881 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5941 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6001 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6061 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6121 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6181 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6241 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6301 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6361 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6421 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6481 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6541 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
