LOCUS Exported 8012 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 HTR2C-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8012) 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 8012) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8012 /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 2567..3568 /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(2634..2653) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3606..3623) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3612..3836 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3882..4310 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3969..3988) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4179..4200 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4319..4339) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4324..4653 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4504..4639 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4566..4585 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4720..5514 /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(4774..4793) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5384..5403 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5688..5809 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5725..5744) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5779..5798 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5858..5874) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5858..5874) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5871..5893) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5882..5898 /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(5906..5936) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5951..5972 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6089..6106) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6260..6845) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6340..6359) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7016..7876) /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 7639..7658 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7877..7981) /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 atatggttaa 1081 tcttcgaaac gcggtccatt cattcctcgt ccacctgatt ggtctcctgg tgtggcagtg 1141 tgatatcagt gtttcccccg tggctgctat cgttaccgat atctttaaca cttctgacgg 1201 tggaagattc aaattccctg acggagtgca gaactggccc gctcttagta ttgtcatcat 1261 tatcattatg acgatcggag gtaacatcct ggtcattatg gcagtgtcaa tggaaaaaaa 1321 acttcacaat gcgacaaact atttcctgat gtccctcgcc atcgccgaca tgctcgtggg 1381 actgctggtc atgccgctgt ccctcctggc aatcctgtac gattacgtgt ggccgttgcc 1441 ccgctatctg tgcccggtct ggatatccct ggacgtgttg ttttctaccg caagtatcat 1501 gcacctgtgt gcaattagcc tggatagata cgtggcaatt agaaacccta ttgaacacag 1561 caggttcaac agcagaacca aagccatcat gaagatcgcg attgtgtggg ccatatctat 1621 cggtgtctca gtgcccatcc ccgtaatcgg cctgagggac gaagagaaag tctttgttaa 1681 caacacaacc tgcgtgttga acgaccccaa cttcgtgctg attggatctt tcgtggcctt 1741 ttttattccg ctcacaatta tggtgattac ctactgcctc acaatatacg ttttgaggcg 1801 acaggccctc atgcttctcc atgggcacac cgaggagccc cccgggctct ctctggattt 1861 ccttaagtgt tgcaagcgga acacagcaga agaggagaac agcgccaacc ctaaccagga 1921 ccaaaacgcc cgccgacgga aaaagaagga gagacgcccc aggggcacta tgcaggctat 1981 caataacgag cgaaaagcat ctaaagtttt gggaattgtg ttttttgtgt ttctgattat 2041 gtggtgcccg ttcttcatca ccaacatcct tagcgttctc tgtgagaaga gctgtaatca 2101 gaagctcatg gagaagctcc tcaacgtgtt tgtctggatt ggttacgttt gcagcgggat 2161 caaccctctg gtgtacaccc tcttcaacaa aatttacaga cgggcattta gcaattattt 2221 gaggtgcaac tacaaagtgg aaaaaaaacc ccctgtgcgc cagatcccaa gagtcgccgc 2281 gaccgctttg tctggcagag agctcaatgt gaacatctac cggcacacaa acgaaccagt 2341 cattgaaaaa gcctcagata atgaaccagg gatcgaaatg caggtggaga atttggaact 2401 gccagtgaac ccctctagcg tggttagtga gcggatatca tcagtgatcg ataccggtgg 2461 acgcacccca cccagcctgg gtccccaaga tgagtcctgc accaccgcca gctcctccct 2521 ggccaaggac acttcatcga ccggtgagaa cctgtacttc cagctaagat tagataaaag 2581 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac 2641 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa 2701 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt 2761 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag 2821 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga 2881 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact 2941 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga 3001 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc 3061 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt 3121 attcggcctt gaattgatca tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc 3181 cgcgtacagc cgcgcgcgta cgaaaaacaa ttacgggtct accatcgagg gcctgctcga 3241 tctcccggac gacgacgccc ccgaagaggc ggggctggcg gctccgcgcc tgtcctttct 3301 ccccgcggga cacacgcgca gactgtcgac ggcccccccg accgatgtca gcctggggga 3361 cgagctccac ttagacggcg aggacgtggc gatggcgcat gccgacgcgc tagacgattt 3421 cgatctggac atgttggggg acggggattc cccgggtccg ggatttaccc cccacgactc 3481 cgccccctac ggcgctctgg atatggccga cttcgagttt gagcagatgt ttaccgatgc 3541 ccttggaatt gacgagtacg gtgggtagct cgagtctaga gggcccgttt aaacccgctg 3601 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 3661 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 3721 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 3781 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 3841 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3901 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3961 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 4021 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 4081 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 4141 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 4201 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 4261 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 4321 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 4381 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 4441 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 4501 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4561 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 4621 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4681 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4741 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 4801 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 4861 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 4921 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 4981 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 5041 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 5101 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 5161 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 5221 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 5281 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 5341 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 5401 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 5461 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 5521 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 5581 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 5641 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 5701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 5821 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 5881 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 5941 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 6001 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6061 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 6601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 6661 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 6721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 6781 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 6841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 6901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 6961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 7021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 7081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 7141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 7201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 7261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 7801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 7861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 7921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 7981 gcacatttcc ccgaaaagtg ccacctgacg tc //