LOCUS Exported 7083 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses strongly red-shifted AlACR1 in mammalian cells. ACCESSION . VERSION . KEYWORDS AlACR1_EYFP_pcDNA3.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7083) AUTHORS Govorunova EG, Sineshchekov OA, Li H, Wang Y, Brown LS, Spudich JL TITLE RubyACRs, nonalgal anion channelrhodopsins with highly red-shifted absorption. JOURNAL Proc Natl Acad Sci U S A. 2020 Sep 15;117(37):22833-22840. doi: 10.1073/pnas.2005981117. Epub 2020 Sep 1. PUBMED 32873643 REFERENCE 2 (bases 1 to 7083) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7083 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 172..551 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 552..755 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 706..726 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 800..819 /label=T7 /note="T7 promoter, forward primer" promoter 800..818 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1772..2488 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1814..1835) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2075..2094) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2422..2443 /label=EGFP-C /note="EGFP, forward primer" CDS 2518..2559 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2569..2586 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2609..2626) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2615..2839 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2885..3313 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2972..2991) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3182..3203 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3322..3342) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3327..3656 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3507..3642 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3569..3588 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3723..4517 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3777..3796) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4387..4406 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4693..4814 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4730..4749) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4784..4803 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4863..4879) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4863..4879) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4876..4898) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4887..4903 /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(4911..4941) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4956..4977 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5094..5111) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5265..5853) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5345..5364) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6024..6884) /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 6647..6666 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6885..6989) /gene="bla" /label=AmpR promoter primer_bind complement(7064..7083) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta 121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat 181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact 601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa 661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta 721 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg 781 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct ggctagttaa 841 gcttggtacc gagctcggat ccatggaggc catggccatg agcgagcacg agctgtggag 901 gagagagttt atcagggtgc tgcagaccca gcccaccaca gacgagacag tgtctcaggg 961 agtggcagga gcaagcctgt gcgtgttcag ctgttccacc gccctgatct gcttttacgg 1021 cttccagtat aggaacggca gatgtaatct gcaccccatc tacgtgcagt ttgtgacatt 1081 ctgcgtgtat tctctggcct gttttggcac cacagatttc ctgtacatcc ctatcgagaa 1141 ccacgccggc gacgtgtttg atctggccat cggcagatat ctgttctgga tgatgacctg 1201 ccctgtgatc atcagcaacg tgaatatcct gctgaatctg atgaccccag acgatctgga 1261 gctgggccac gtgaccttca tgatggtgaa ggacatcgtg atgatcgcct tcggcgtgct 1321 ggccgccatg cagaagaacg tgaccctgaa gtggatcttt aatctggcct cctacttcgt 1381 gtgcttctgg ctggtggtgg atctgctggc catgatccgg gagaagaaga agcacttcat 1441 cgtgcacgag aggtgctggg agtgggtgct ggccgtgatg acatacttct ttttcgccca 1501 gagcctgttt atcttcctgt atgcctttgg ccccccttgt ttcaacctga tgggcccaga 1561 cggcgataag atcggccact ccgtgggcga cctgtttgcc aagaatctgt tcggcttttt 1621 cgcctggtac gtgaggtttg tggtgctgga gcccttcgtg aagaggatca ggagcgccag 1681 ggcagtgacc ccatatcaga ggggctctga tgcaatggca ccacacaaga tgaggaacgt 1741 gtctttcctg aatagaaaga cagcggccgc cgtgagcaag ggcgaggagc tgttcaccgg 1801 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc 1861 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac 1921 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg 1981 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga 2041 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc 2101 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt 2161 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt 2221 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa 2281 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga 2341 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga 2401 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 2461 tctcggcatg gacgagctgt acaagtaaca tatgctctag agggcccgcg gttcgaaggt 2521 aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca tcatcaccat 2581 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 2641 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 2701 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 2761 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 2821 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 2881 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2941 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3001 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 3061 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3121 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3181 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3241 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3301 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3481 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3541 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3601 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3661 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3721 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3781 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3841 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3901 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3961 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4021 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4081 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4141 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4201 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4261 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4321 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4381 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 4441 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 4501 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 4561 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4621 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4681 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4741 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 4801 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 4861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 4921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 4981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cctttttcca taggctccgc 5281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 5461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 5761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 5821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 5881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 5941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 6781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 6841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 6901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 6961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7021 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 7081 ccg //