LOCUS       Exported                5780 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  optogenetic p38 inhibitor with RET to LOV2 to increase sensitivity.
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Synapsin-mTurquoise2-optop38i3-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5780)
  AUTHORS   Li LL, Klein FM, Li Greci L, Popinigis A, Freudenberg F, Courtney MJ
  TITLE     Resonance energy transfer sensitises and monitors in situ switching 
            of LOV2-based optogenetic actuators.
  JOURNAL   Nat Commun. 2020 Oct 9;11(1):5107. doi: 10.1038/s41467-020-18816-8.
  PUBMED    33037199
REFERENCE   2  (bases 1 to 5780)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5780
                     /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        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      652..661
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             658..1374
                     /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"
     primer_bind     complement(703..724)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(964..983)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1311..1332
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1909..2497
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1962..1982)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2380..2391)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2527..3003
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2726..2745)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3043..3183
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3043..3172
                     /label=AAV2 ITR
     rep_origin      3258..3713
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3345..3364)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3555..3576
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3730..3749)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3849..3871
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3909..3927)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3995..4099
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4100..4960
                     /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(4318..4337)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5131..5719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5620..5639
                     /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 gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtacagctag cgctaccggt cgccaccatg
      661 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      721 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      781 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      841 gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      901 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      961 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1021 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1081 ctggagtaca actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc
     1141 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
     1201 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1261 ctgagcaccc agtccaagct tagcaaagac cccaacgaga agcgcgatca catggtcctg
     1321 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1381 ctcagatctc gagctcaagc ttcgaattca ctggcaacaa cactggagag aatcgagaaa
     1441 aacttcgtca tcaccgaccc tagactgccc gataatccta tcatcttcgc ctccgactca
     1501 tttctccagc tcactgagta ctctcgggag gaaattctgg gccggaattg ccggttcctc
     1561 cagggacctg aaaccgatag agccactgtc cgaaaaatcc gggatgccat cgacaatcag
     1621 actgaggtga ccgtccagct catcaattac accaaatctg gcaaaaaatt ctggaacctg
     1681 ttccacctcc agcctatgag agatcagaag ggcgacgtcc agtactttat tggcgtccag
     1741 ctggatggaa ctgaacatgt gagggacgct gcagaacgag agggcgtgat gctcatcaaa
     1801 aaaaccgccg aaaacatcga tgaggccgct aaagagctgt ctagaaagcg gaagaaggac
     1861 ctgcggatct cctgcatgtt ctaggatccg cggccgctag ataactgaaa tcaacctctg
     1921 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1981 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggccttcatt
     2041 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     2101 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2161 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2221 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2281 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc
     2341 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2401 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2461 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gattgtaccg cgggcccggg
     2521 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     2581 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2641 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     2701 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     2761 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     2821 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     2881 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     2941 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3001 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     3061 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3121 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3181 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3241 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3301 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3361 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3421 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3481 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3541 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3601 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3661 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3721 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3781 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3841 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3901 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3961 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4021 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4081 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4141 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4201 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4261 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4321 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4381 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4441 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4501 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4561 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4621 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4681 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4741 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4801 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4861 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4921 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5761 ctggcctttt gctcacatgt
//
