LOCUS Exported 7715 bp ds-DNA circular SYN 05-OCT-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 CXCR2-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7715) 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 7715) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 5, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7715 /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 2039..3040 /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(2106..2125) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3072..3089) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3078..3302 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3348..3776 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3435..3454) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3645..3666 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3785..3805) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3790..4119 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3970..4105 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4032..4051 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4186..4980 /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(4240..4259) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4850..4869 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5154..5275 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5191..5210) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5245..5264 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5324..5340) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5324..5340) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5337..5359) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5348..5364 /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(5372..5402) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5417..5438 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5555..5572) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5726..6314) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5806..5825) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6485..7345) /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 7108..7127 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7346..7450) /gene="bla" /label=AmpR promoter primer_bind complement(7525..7544) /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 ggaggacttc aacatggagt cagactcctt cgaagacttc tggaaaggtg aggacctgtc 901 aaactattct tatagtagca cactccctcc atttcttttg gacgctgcac cctgcgagcc 961 agaatccctg gagattaata agtactttgt ggttatcata tacgcactcg tgtttttgct 1021 gtcactgctc gggaacagcc ttgtcatgct tgtgattctc tactctcggg tgggccggtc 1081 agttaccgat gtgtatcttc tgaacctggc acttgcggat ttgctcttcg ctctcactct 1141 ccccatctgg gcagcctcca aggttaacgg ctggatcttt ggcaccttcc tttgcaaggt 1201 cgtctcactg ctgaaagaag tgaatttcta ctcaggcatt ctgctgcttg cctgcatcag 1261 cgtggacaga tatctggcga tcgtgcacgc cacacggaca ctgactcaaa agaggtatct 1321 cgtgaaattc atatgccttt ccatttgggg gctgtcactg ctgctcgcct tgcctgtctt 1381 gttgtttaga cggaccgtat atagctccaa tgtgagcccc gcatgctatg aggatatggg 1441 caataataca gccaactggc gaatgctgct cagaatcctg ccccagtcat tcggatttat 1501 cgttcccctg ctcattatgc ttttctgtta cgggtttact ctgcggaccc ttttcaaagc 1561 tcatatgggc cagaagcaca gagcaatgcg ggttatcttc gctgtggtcc tgatatttct 1621 gctgtgttgg ctgccctaca acctcgtcct gctcgccgac actctgatgc ggactcaggt 1681 gattcaggag acatgcgagc ggcgcaacca catcgacaga gccctggacg ctacagaaat 1741 acttggcatt ctccattctt gtttgaaccc actgatatac gcctttatag gacaaaagtt 1801 taggcacggg ctgctgaaaa tcctcgccat ccatggcctt atctccaagg attccctccc 1861 caaggactcc cgcccaagct tcgtgggttc atcctccggc cacacctcca cgaccctgat 1921 cgataccggt ggacgcaccc cacccagcct gggtccccaa gatgagtcct gcaccaccgc 1981 cagctcctcc ctggccaagg acacttcatc gaccggtgag aacctgtact tccagctaag 2041 attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga 2101 aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg 2161 gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca 2221 ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc 2281 taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac 2341 acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca 2401 aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg 2461 ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac 2521 tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga 2581 gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg 2641 tgaaagtggg tccgcgtaca gccgcgcgcg tacgaaaaac aattacgggt ctaccatcga 2701 gggcctgctc gatctcccgg acgacgacgc ccccgaagag gcggggctgg cggctccgcg 2761 cctgtccttt ctccccgcgg gacacacgcg cagactgtcg acggcccccc cgaccgatgt 2821 cagcctgggg gacgagctcc acttagacgg cgaggacgtg gcgatggcgc atgccgacgc 2881 gctagacgat ttcgatctgg acatgttggg ggacggggat tccccgggtc cgggatttac 2941 cccccacgac tccgccccct acggcgctct ggatatggcc gacttcgagt ttgagcagat 3001 gtttaccgat gcccttggaa ttgacgagta cggtgggtag tctagagggc ccgtttaaac 3061 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc 3121 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga 3181 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga 3241 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat 3301 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag 3361 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 3421 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 3481 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 3541 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 3601 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 3661 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 3721 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 3781 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 3841 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 3961 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 4021 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 4081 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 4141 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 4201 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 4261 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 4321 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 4381 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 4441 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 4501 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 4561 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 4621 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 4681 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 4741 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 4801 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 4861 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 4921 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 4981 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 5041 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 5101 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt 5161 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag 5221 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg 5281 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 5341 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 5401 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 5461 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 5521 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 5581 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 5641 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 5701 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 5761 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 5821 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 5881 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 5941 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6001 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 6061 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 6121 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 6181 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 6241 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 6301 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 6361 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 6421 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 6481 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 6541 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 6601 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 6661 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 6721 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 6781 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 6841 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 6901 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 6961 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 7021 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 7081 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 7141 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 7201 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 7261 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 7321 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 7381 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 7441 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc 7501 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta 7561 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac 7621 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc 7681 gctgcttcgc gatgtacggg ccagatatac gcgtt //