LOCUS Exported 5957 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses NES-CRY2PHR-TevC of BLITz system. ACCESSION . VERSION . KEYWORDS pCMV-NES-CRY2PHR-TevC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5957) AUTHORS Lee D, Creed M, Jung K, Stefanelli T, Wendler DJ, Oh WC, Mignocchi NL, Luscher C, Kwon HB TITLE Temporally precise labeling and control of neuromodulatory circuits in the mammalian brain. JOURNAL Nat Methods. 2017 May;14(5):495-503. doi: 10.1038/nmeth.4234. Epub 2017 Apr 3. PUBMED 28369042 REFERENCE 2 (bases 1 to 5957) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5957 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..978 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" promoter 1013..1031 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1013..1030 /label=SP6 /note="SP6 promoter, forward primer" CDS 1072..1098 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 1108..2601 /codon_start=1 /product="N-terminal photolyase homology region of Arabidopsis thaliana cryptochrome 2" /label=CRY2PHR /note="binds to CIB1 upon exposure to blue light (Kennedy et al., 2010)" /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA" primer_bind complement(2964..2983) /label=T7 /note="T7 promoter, forward primer" promoter complement(2965..2983) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2988..3122 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3012..3031) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3066..3085 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3235..3255) /label=T3 /note="T3 promoter, forward primer" promoter complement(3235..3253) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3274..3290) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3274..3290) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3287..3309) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3298..3314 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3322..3352) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3367..3388 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3505..3522) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3675..4263) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3755..3774) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4434..5294) /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 5057..5076 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5295..5399) /gene="bla" /label=AmpR promoter rep_origin complement(5425..5880) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5562..5583) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5774..5793 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt 61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca 121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt 181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg 241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat 301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca 361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt 421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac 481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac 541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta 601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa 661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg 721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac 781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg 841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa 901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc 961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt 1021 gacactatag aatacaagct acttgttctt tttgcaggat ccatgcttca acttcctcct 1081 cttgaacgtc ttactctcga ggccaccatg aagatggaca aaaagactat agtttggttt 1141 agaagagacc taaggattga ggataatcct gcattagcag cagctgctca cgaaggatct 1201 gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc tggaagagct 1261 tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa ggctcttgga 1321 tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga ttgtatccgc 1381 gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc gttagttcgg 1441 gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag ctacaatgga 1501 gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt tacgagtttc 1561 aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct tcctcctcct 1621 tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc gattgaagaa 1681 ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag agcttggtct 1741 ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca gttgatagat 1801 tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc tccgtatctc 1861 catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa acaaattata 1921 tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct taggggaatc 1981 ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca cgagcaatcg 2041 ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt caaggcctgg 2101 agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct ttgggctacc 2161 ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa gtttcttctc 2221 cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc tgatttggaa 2281 tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca cgagcttgat 2341 cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga gtacataagg 2401 caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc atgggacgct 2461 cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc gaaacccatt 2521 gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac ccgtgaagca 2581 cagatcatga tcggagcagc actgcaggag ctcggtggag gcggccgctc cggtggcggt 2641 ggctctacta agagcatgtc cagcatggtg agcgatacta gctgtacctt cccatcatct 2701 gacggaatct tctggaagca ctggattcag actaaggacg gccagtgtgg cagcccactg 2761 gtgagcacac gagacggatt catcgtgggg attcacagcg cctccaactt tacaaacacc 2821 aataactatt tcacctcagt gccaaagaac tttatggagc tgctgaccaa ccaggaggcc 2881 cagcagtggg tgagcgggtg gcgcctgaac gccgattccg tgctgtgggg cgggcacaag 2941 gtgtttatgg tgtgagctag cctccctata gtgagtcgta ttacgtagat ccagacatga 3001 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta 3061 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag 3121 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt 3181 tttaattcgc gtccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta 3241 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg 3301 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg 3361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 3421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 3481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 3541 gcgggagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 3781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 3841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 3901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 3961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 4141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 4381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 4441 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 4501 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 4561 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 4621 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 4681 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 4741 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 4801 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 4861 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 4921 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 4981 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 5041 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 5101 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 5161 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 5221 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 5281 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5401 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt 5461 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt 5521 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 5581 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 5641 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac 5701 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 5761 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 5821 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt 5881 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 5941 tattacgcca gtcgacc //