LOCUS Exported 6632 bp ds-DNA linear SYN 01-JUN-2021 DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel receptorome expression and screening via transcriptional output, with transcriptional activation following arrestin translocation. ACCESSION . VERSION . KEYWORDS GPR61-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6632) AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL TITLE PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20. PUBMED 25895059 REFERENCE 2 (bases 1 to 6632) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 1, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6632 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1187..2188 /codon_start=1 /product="tetracycline-controlled transactivator, comprising a fusion of the tetracycline repressor TetR with the C-terminal activation domain of herpes simplex virus VP16" /label=tTA /note="In the Tet-Off(R) system, tTA binds to the Tet-responsive element and stimulates transcription only in the absence of tetracycline or doxycycline." /translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIEG LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADA LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG" primer_bind complement(1254..1273) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(2226..2243) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2232..2456 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2502..2930 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2589..2608) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2799..2820 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2939..2959) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2944..3273 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3124..3259 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3186..3205 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3340..4134 /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(3394..3413) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4004..4023 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4308..4429 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4345..4364) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4399..4418 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4478..4494) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4478..4494) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4491..4513) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4502..4518 /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(4526..4556) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4571..4592 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4709..4726) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4880..5465) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4960..4979) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5636..6496) /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 6259..6278 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6497..6601) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc 961 agatatccag cacagtggcg gccgcgccac catgaagacg atcatcgccc tgagctacat 1021 cttctgcctg gtattcgccg actacaagga cgatgatgac gccagcatcg ataccggtgg 1081 acgcacccca cccagcctgg gtccccaaga tgagtcctgc accaccgcca gctcctccct 1141 ggccaaggac acttcatcga ccggtgagaa cctgtacttc cagctaagat tagataaaag 1201 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac 1261 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa 1321 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt 1381 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag 1441 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga 1501 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact 1561 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga 1621 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc 1681 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt 1741 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc 1801 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga 1861 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct 1921 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga 1981 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt 2041 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc 2101 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc 2161 ccttggaatt gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg 2221 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 2281 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 2341 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 2401 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 2461 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 2521 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 2581 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 2641 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 2701 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 2761 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 2821 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 2881 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 2941 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3001 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3061 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 3121 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 3181 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 3241 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 3301 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 3361 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 3421 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 3481 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 3541 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 3601 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 3661 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 3721 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 3781 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 3841 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 3901 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 3961 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 4021 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4081 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 4141 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 4201 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 4261 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 4321 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 4381 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 4441 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 4501 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 4561 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 4621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 4681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 4741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 4801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 4861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 4921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 4981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 5041 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 5101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 5161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 5221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 5281 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 5341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 5401 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 5461 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 5521 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 5581 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 5641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 5701 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 5761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 5821 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 5881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 5941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 6001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 6061 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 6121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 6181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 6241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 6301 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 6361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 6421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 6481 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 6541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 6601 gcacatttcc ccgaaaagtg ccacctgacg tc //