LOCUS Exported 7688 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 FPR2-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7688) 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 7688) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7688 /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 2246..3247 /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(2313..2332) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3279..3296) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3285..3509 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3555..3983 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3642..3661) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3852..3873 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3992..4012) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3997..4326 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4177..4312 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4239..4258 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4393..5187 /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(4447..4466) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5057..5076 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5361..5482 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5398..5417) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5452..5471 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5531..5547) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5531..5547) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5544..5566) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5555..5571 /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(5579..5609) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5624..5645 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5762..5779) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5933..6521) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6013..6032) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6692..7552) /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 7315..7334 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7553..7657) /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 atatggaaac 1081 caatttctct actcccctga atgagtacga agaggtgtct tacgaatcag cgggctacac 1141 agtcctgcga atcctcccgc ttgtcgtact tggcgtgact tttgtgctcg gtgttcttgg 1201 taacgggttg gtaatctggg tggcagggtt cagaatgact cgcacggtga ctaccatttg 1261 ctacctcaac ctggccctgg ccgatttttc ctttactgct actctcccgt ttctcattgt 1321 gtctatggcc atgggcgaga aatggccttt cgggtggttc ctgtgcaagc tcatccacat 1381 tgtggtggac atcaatcttt ttggtagtgt cttcctgata ggctttattg cactcgatag 1441 gtgtatttgt gtactgcacc cagtctgggc ccagaatcac agaaccgtca gccttgctat 1501 gaaagtgata gttggtccat ggatcttggc cctggtgctt accctgccag ttttcttgtt 1561 tctcaccacc gtgaccatcc caaatggcga cacatactgc acatttaatt tcgcaagctg 1621 ggggggtacg cctgaggagc gattgaaggt tgcgataacc atgttgacag ccagaggaat 1681 aatcagattc gtgattggct tctccctgcc gatgtctata gtggctatct gctacggcct 1741 catcgcggct aagatccata agaaaggtat gatcaagtct tccaggcctc tccgggtatt 1801 gacagccgta gttgcaagtt tctttatatg ctggttccct ttccagctgg tggcactgct 1861 gggaactgtg tggctgaaag aaatgttgtt ctatgggaag tataaaataa ttgacattct 1921 cgtaaaccct accagcagtc tggctttttt taactcttgc ctgaacccaa tgctgtatgt 1981 ttttgtgggc caggacttcc gagaacggct gattcacagc cttcctacct cactggagag 2041 agccctgagc gaagatagtg cgcccacgaa cgacaccgcc gccaactccg cttctcctcc 2101 tgctgagaca gaactccaag cgatgatcga taccggtgga cgcaccccac ccagcctggg 2161 tccccaagat gagtcctgca ccaccgccag ctcctccctg gccaaggaca cttcatcgac 2221 cggtgagaac ctgtacttcc agctaagatt agataaaagt aaagtgatta acagcgcatt 2281 agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct 2341 aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc 2401 cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag 2461 ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg 2521 cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga 2581 aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact 2641 cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc 2701 taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac gacaagctat 2761 cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat 2821 atgcggatta gaaaaacaac ttaaatgtga aagtgggtcc gcgtacagcc gcgcgcgtac 2881 gaaaaacaat tacgggtcta ccatcgaggg cctgctcgat ctcccggacg acgacgcccc 2941 cgaagaggcg gggctggcgg ctccgcgcct gtcctttctc cccgcgggac acacgcgcag 3001 actgtcgacg gcccccccga ccgatgtcag cctgggggac gagctccact tagacggcga 3061 ggacgtggcg atggcgcatg ccgacgcgct agacgatttc gatctggaca tgttggggga 3121 cggggattcc ccgggtccgg gatttacccc ccacgactcc gccccctacg gcgctctgga 3181 tatggccgac ttcgagtttg agcagatgtt taccgatgcc cttggaattg acgagtacgg 3241 tgggtagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg 3301 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 3361 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 3421 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 3481 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 3541 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 3601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 3661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 3721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 3781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 3841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 3901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 3961 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 4021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 4081 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 4141 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 4201 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 4261 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 4321 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 4381 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 4441 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 4501 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 4561 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 4621 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 4681 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 4741 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 4801 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 4861 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 4921 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 4981 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 5041 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 5101 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 5161 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 5221 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 5281 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 5341 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 5401 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 5461 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 5521 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 5581 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 5641 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 5701 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 5761 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 5821 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 5881 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 5941 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 6001 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 6061 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 6121 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 6181 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 6241 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 6301 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 6361 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 6421 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 6481 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 6541 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 6601 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 6661 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 6721 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 6781 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 6841 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 6901 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 6961 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 7021 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 7081 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 7141 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 7201 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 7261 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 7321 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 7381 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 7441 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 7501 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 7561 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 7621 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 7681 ctgacgtc //