LOCUS Exported 6211 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Reporter construct. ACCESSION . VERSION . KEYWORDS TRE-ChrimsonR-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6211) AUTHORS Wang W, Wildes CP, Pattarabanjird T, Sanchez MI, Glober GF, Matthews GA, Tye KM, Ting AY TITLE A light- and calcium-gated transcription factor for imaging and manipulating activated neurons. JOURNAL Nat Biotechnol. 2017 Jun 26. doi: 10.1038/nbt.3909. PUBMED 28650461 REFERENCE 2 (bases 1 to 6211) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6211 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 77..95 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 112..130 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 148..166 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 184..202 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 219..237 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 255..273 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" promoter 285..323 /label=minimal CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 320..344 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 372..381 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 378..1427 /codon_start=1 /product="red light-drivable channelrhodopsin from Chlamydomonas noctigama (Klapoetke et al., 2014)" /label=ChrimsonR /note="K176R mutant with accelerated closing kinetics" /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE FVEEEDEDTV" regulatory 1491..1500 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1497..2207 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1642..1660) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1910..1929 /label=mCherry-F /note="mCherry, forward primer" misc_feature 2239..2827 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2292..2312) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2710..2721) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2859..3335 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3058..3077) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3375..3515 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3375..3504 /label=AAV2 ITR rep_origin 3590..4045 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3677..3696) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3887..3908 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4062..4081) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4181..4203 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4241..4259) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4327..4431 /gene="bla" /label=AmpR promoter CDS 4432..5292 /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(4650..4669) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5463..6051 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5952..5971 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 6113..31 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" ORIGIN 1 ggagtggcca actccatcac taggggttcc tgcggccgca cgcgtccttc aggtccgagg 61 ttctagacga gtttactccc tatcagtgat agagaacgat gtcgagttta ctccctatca 121 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt 181 tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg atagagaacg 241 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg cgtgtacggt 301 gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcaaag ggcgaattcg 361 atccaccggt cgccaccatg gctgagctga tcagcagcgc caccagatct ctgtttgccg 421 ccggaggcat caacccttgg cctaacccct accaccacga ggacatgggc tgtggaggaa 481 tgacacctac aggcgagtgc ttcagcaccg agtggtggtg tgacccttct tacggactga 541 gcgacgccgg atacggatat tgcttcgtgg aggccacagg cggctacctg gtcgtgggag 601 tggagaagaa gcaggcttgg ctgcacagca gaggcacacc aggagaaaag atcggcgccc 661 aggtctgcca gtggattgct ttcagcatcg ccatcgccct gctgacattc tacggcttca 721 gcgcctggaa ggccacttgc ggttgggagg aggtctacgt ctgttgcgtc gaggtgctgt 781 tcgtgaccct ggagatcttc aaggagttca gcagccccgc cacagtgtac ctgtctaccg 841 gcaaccacgc ctattgcctg cgctacttcg agtggctgct gtcttgcccc gtgatcctga 901 tcagactgag caacctgagc ggcctgaaga acgactacag caagcggacc atgggcctga 961 tcgtgtcttg cgtgggaatg atcgtgttcg gcatggccgc aggactggct accgattggc 1021 tcaagtggct gctgtatatc gtgtcttgca tctacggcgg ctacatgtac ttccaggccg 1081 ccaagtgcta cgtggaagcc aaccacagcg tgcctaaagg ccattgccgc atggtcgtga 1141 agctgatggc ctacgcttac ttcgcctctt ggggcagcta cccaatcctc tgggcagtgg 1201 gaccagaagg actgctgaag ctgagccctt acgccaacag catcggccac agcatctgcg 1261 acatcatcgc caaggagttt tggaccttcc tggcccacca cctgaggatc aagatccacg 1321 agcacatcct gatccacggc gacatccgga agaccaccaa gatggagatc ggaggcgagg 1381 aggtggaagt ggaagagttc gtggaggagg aggacgagga cacagtggcg gcaccggtag 1441 tagcagtgag cggtggaagc ggttctggaa gtgctcggga tccaccggtc gccaccatgg 1501 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc 1561 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 1621 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg 1681 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg 1741 ccgacatccc cgactacttg aagctgtcct tccccgaggg attcaagtgg gagcgcgtga 1801 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg 1861 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc 1921 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc 1981 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg 2041 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca 2101 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg 2161 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaact agtaccggta 2221 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt 2281 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 2341 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 2401 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 2461 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 2521 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2581 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 2641 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc 2701 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2761 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 2821 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct 2881 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa 2941 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga 3001 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct 3061 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct 3121 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga 3181 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc 3241 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat 3301 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg 3361 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 3421 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 3481 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 3541 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 3601 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3661 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3721 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 3781 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 3841 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 3901 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg 3961 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 4021 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 4081 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 4141 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 4201 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 4261 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 4321 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 4381 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 4441 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 4501 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 4561 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 4621 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 4681 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 4741 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 4801 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 4861 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 4921 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 4981 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 5041 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 5101 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 5161 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 5221 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 5281 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 5341 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5401 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5461 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5521 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5581 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5641 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5701 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5761 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5821 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5881 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5941 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 6001 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 6061 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg 6121 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc 6181 ggcctcagtg agcgagcgag cgcgcagaga g //