LOCUS Exported 8561 bp ds-DNA circular SYN 12-MAY-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 GPR111-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8561) 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 8561) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8561 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2885..3886 /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(2952..2971) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3918..3935) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3924..4148 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4194..4622 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4281..4300) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4491..4512 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4631..4651) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4636..4965 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4816..4951 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4878..4897 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5032..5826 /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(5086..5105) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5696..5715 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6000..6121 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6037..6056) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6091..6110 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6170..6186) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6170..6186) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6183..6205) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6194..6210 /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(6218..6248) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6263..6284 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6401..6418) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6572..7160) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6652..6671) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7331..8191) /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 7954..7973 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8192..8296) /gene="bla" /label=AmpR promoter primer_bind complement(8371..8390) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg 661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa 721 ttctgcagat atccagcaca gtggcggccg cgccaccatg aagacgatca tcgccctgag 781 ctacatcttc tgcctggtat tcgccgacta caaggacgat gatgacgcca gcatcgatat 841 gacacatatc cttctgttgt attatctggt gtttctgctg cctaccgagt catgcaggac 901 tctgtaccaa gcggcaagta agtccaaaga gaaggtcccc gcacgcccac acggagtttg 961 cgacggtgtg tgtacagact acagccagtg tacacagcca tgcccccctg acacacaggg 1021 aaacatgggg tttagctgtc ggcaaaaaac atggcataag atcaccgaca cttgccagac 1081 cctgaacgct ttgaacatat tcgaagaaga tagtaggctg gtgcaaccct tcgaggataa 1141 tataaaaatc tctgtttaca ctggcaaatc cgagacaatt actgatatgc tgcttcaaaa 1201 gtgtcctacg gacttgagtt gcgtgattcg caacattcaa cagagtccct ggataccagg 1261 caatatcgcc gttattgtgc aacttttgca taacattagt acggccatct ggaccggcgt 1321 cgacgaagcg aaaatgcagt cctatagtac gatcgcgaat catatcctga attctaaaag 1381 catctccaat tggacattta tccctgatcg gaactcctca tatatcttgc tgcacagcgt 1441 caattcattc gcaaggagac tttttattga taaacatcct gtcgatatct cagatgtttt 1501 catccacact atgggtacca ctatttcagg tgacaacatc gggaagaact tcactttttc 1561 aatgcgcatc aacgacacat ctaatgaagt aactggccgc gtattgatta gccgagatga 1621 actcaggaag gtcccctctc catctcaggt tatttccatc gcgtttccaa ccattggagc 1681 aatactggaa gcatctctcc ttgaaaatgt taccgtcaat ggtctggtcc tttctgccat 1741 actgccaaaa gagctgaaaa gaatttccct catctttgaa aagattagta aatcagagga 1801 aagacgaact cagtgcgtag gttggcactc agtggagaat aggtgggatc aacaggcatg 1861 taagatgata caggaaaaca gccagcaagc agtgtgcaaa tgtcgcccct ctaaactctt 1921 cacctctttc tccatcctga tgtctcctca catactggag tccctcatct tgacttacat 1981 tacatatgtg ggcctgggga tttctatatg ttccctgatc ctctgtcttt ctattgaagt 2041 tcttgtttgg tcacaggtta caaagacgga gattacttac ctgagacatg tttgcatagt 2101 caacatcgca gccacattgc tgatggctga tgtatggttt atcgtggcca gttttctgtc 2161 tggacctata acacaccaca aaggatgcgt tgccgcaacc ttcttcgttc acttctttta 2221 tctgtctgtt tttttttgga tgcttgccaa agccctgttg atcctctacg gaataatgat 2281 cgtattccac accctcccta aaagcgtact tgtggcctct ctgttttcag tgggctacgg 2341 ttgtcccctg gccatagctg caatcaccgt ggcggccaca gaaccaggga aaggatatct 2401 ccggccagaa atctgctggc tgaactggga catgactaag gccctgcttg cctttgtcat 2461 ccctgcgctt gctatagtgg tagtcaactt gatcacggtg acactggtga tagtaaaaac 2521 gcagcgggct gctatcggta atagcatgtt ccaggaagta agagctattg tacggatcag 2581 caaaaatatc gcgatcctga cccccctcct gggattgacc tggggattcg gcgtagccac 2641 cgtcatagac gacagaagct tggcctttca tatcattttc agcttgctga atgcattcca 2701 aggctttttt atcctggtct ttggaacaat tctggacccg aagatcagag aggctctgaa 2761 ggggatcgat accggtggac gcaccccacc cagcctgggt ccccaagatg agtcctgcac 2821 caccgccagc tcctccctgg ccaaggacac ttcatcgacc ggtgagaacc tgtacttcca 2881 gctaagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg 2941 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt 3001 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga 3061 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa 3121 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt 3181 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg 3241 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac 3301 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc 3361 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg 3421 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact 3481 taaatgtgaa agtgggtccg cgtacagccg cgcgcgtacg aaaaacaatt acgggtctac 3541 catcgagggc ctgctcgatc tcccggacga cgacgccccc gaagaggcgg ggctggcggc 3601 tccgcgcctg tcctttctcc ccgcgggaca cacgcgcaga ctgtcgacgg cccccccgac 3661 cgatgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc 3721 cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg 3781 atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga 3841 gcagatgttt accgatgccc ttggaattga cgagtacggt gggtagtcta gagggcccgt 3901 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 3961 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 4021 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 4081 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 4141 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 4201 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 4261 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 4321 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 4381 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 4441 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 4501 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 4561 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 4621 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 4681 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 4741 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 4801 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 4861 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 4921 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 4981 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 5041 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 5101 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 5161 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 5221 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 5281 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 5341 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 5401 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 5461 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 5521 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 5581 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 5641 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 5701 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 5761 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 5821 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 5881 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 5941 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 6001 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6061 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6121 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 6181 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 6241 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 6301 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 6361 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 6421 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 6481 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 6541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 6601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 6661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 6721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 6781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 6841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 6901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 6961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 7021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 7081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 7141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 7201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 7261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 7321 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 7381 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 7441 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 7501 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 7561 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 7621 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 7681 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 7741 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 7801 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 7861 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 7921 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 7981 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 8041 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 8101 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 8161 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 8221 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 8281 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 8341 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag 8401 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 8461 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg 8521 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t //