LOCUS Exported 7010 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION fusion protein of Gaussia luciferase variant superluminescent, Leptosphaeria maculans opsin, and enhanced yellow fluorescent protein for bioluminescent optogenetics. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-iLMO (slGLuc-Mac-EYFP) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7010) AUTHORS Berglund K, Clissold K, Li HE, Wen L, Park SY, Gleixner J, Klein ME, Lu D, Barter JW, Rossi MA, Augustine GJ, Yin HH, Hochgeschwender U TITLE Luminopsins integrate opto- and chemogenetics by using physical and biological light sources for opsin activation. JOURNAL Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E358-67. doi: 10.1073/pnas.1510899113. Epub 2016 Jan 5. PUBMED 26733686 REFERENCE 2 (bases 1 to 7010) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7010 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 5..145 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 16..145 /label=AAV2 ITR promoter 190..637 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 1278..1287 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2400..3116 /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 2400..3113 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2442..2463) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2703..2722) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3050..3071 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3141..3729 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3194..3214) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3612..3623) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3761..4237 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3960..3979) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4277..4417 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4277..4406 /label=AAV2 ITR rep_origin 4492..4947 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4579..4598) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4789..4810 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4964..4983) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5083..5105 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5143..5161) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5229..5333 /gene="bla" /label=AmpR promoter CDS 5334..6194 /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(5552..5571) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6365..6953 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6854..6873 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 atgtcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg 61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg 121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc 181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct 241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc 301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc 361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac 421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg 481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg 541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt 601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta 661 gagtcgagct caagcttggc accatgggag tcaaagttct gtttgccctg atctgcatcg 721 ctgtggccga ggccaagccc accgagaaca acgaagactt caacatcgtg gccgtggcca 781 gcaacttcgc gaccacggat ctcgatgctg accgcgggaa gttgcccggc aagaagctgc 841 cgctggaggt gctcaaagag atggaagcca atgcccggaa agctggctgc accaggggct 901 gtctgatctg cctgtcccac atcaagtgca cgcccaagat gaagaagtgg ctgccaggac 961 gctgccacac ctacgaaggc gacaaagagt ccgcacaggg cggcataggc gaggcgatcg 1021 tcgacattcc tgagattcct gggttcaagg acttggagcc catggagcag ttcatcgcac 1081 aggtcgatct gtgtgtggac tgcacaactg gctgcctcaa agggcttgcc aacgtgcagt 1141 gttctgacct gctcaagaag tggctgccgc aacgctgtgc gacctttgcc agcaagatcc 1201 agggccaggt ggacaagatc aagggggccg gtggtgacgg cgacccacta gtccagtgtg 1261 gtggaattgc cggatctgcc accatgggga cctggatgct gacgaaggct cgcgaggctg 1321 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt caatcaagct ggttctccat 1381 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg 1441 ccaccatgat cgtggaccag ttcgaggagg tgctgatgaa gaccagccag ctgttcccac 1501 tgccaaccgc tacccagagc gcccagccaa cccacgtggc ccccgtgcca accgtgctgc 1561 ccgacacccc catctacgag accgtgggcg acagcggcag caagaccctg tgggtggtgt 1621 tcgtgctgat gctgatcgcc agcgccgcct tcaccgccct gagctggaag atccccgtga 1681 acaggaggct gtaccacgtg atcaccacca tcatcaccct gaccgccgcc ctgagctact 1741 tcgctatggc taccggccac ggagtggccc tgaacaagat cgtgatcagg acccagcacg 1801 accacgtgcc cgacacctac gagaccgtgt accgacaggt gtactacgcc aggtacatcg 1861 actgggctat caccacccca ctgctgctgc tggacctggg actgctggct ggaatgagcg 1921 gagcccacat cttcatggcc atcgtggctg acctgatcat ggtgctgacc ggcctgttcg 1981 ctgctttcgg cagcgaggga accccacaga agtggggatg gtacaccatc gcctgcatcg 2041 cctacatctt cgtggtgtgg cacctggtgc tgaacggcgg cgccaacgcc agggtgaagg 2101 gcgagaagct gaggagcttc ttcgtggcca tcggagctta caccctgatc ctgtggaccg 2161 cttacccaat cgtgtgggga ctggctgacg gagctaggaa gatcggagtg gacggagaga 2221 tcatcgctta cgctgtgctg gacgtgctgg ctaagggagt gttcggagct tggctgctgg 2281 tgacccacgc caacctgagg gagagcgacg tggagctgaa cggcttctgg gccaacggcc 2341 tgaacaggga gggcgccatc aggatcggcg aggacgacgg cgcccgaccg gtagtagcag 2401 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 2461 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 2521 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 2581 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 2641 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 2701 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 2761 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 2821 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 2881 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 2941 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 3001 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 3061 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat 3121 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg 3181 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt 3241 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc 3301 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt 3361 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga 3421 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct 3481 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat 3541 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct 3601 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc 3661 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg 3721 cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc cctgtgaccc 3781 ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct 3841 aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg 3901 gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt 3961 ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc 4021 ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat 4081 gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag 4141 gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg 4201 attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg taaccacgtg 4261 cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct 4321 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg 4381 gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc 4441 cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct 4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 4561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 4681 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct 4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 4801 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt 4861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 4921 ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg 4981 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 5041 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 5101 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 5161 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 5221 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 5281 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 5341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 5401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 5461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 5521 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 5581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 5641 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 5701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 5761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 5821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 5881 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 5941 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 6001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 6061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 6121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 6181 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 6241 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 6301 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 6361 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 6421 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 6481 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 6541 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 6601 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 6661 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 6721 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 6781 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 6841 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 6901 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 6961 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac //