LOCUS Exported 7967 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 DRD2-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7967) 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 7967) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7967 /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 2522..3523 /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(2589..2608) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3561..3578) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3567..3791 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3837..4265 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3924..3943) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4134..4155 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4274..4294) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4279..4608 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4459..4594 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4521..4540 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4675..5469 /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(4729..4748) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5339..5358 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5643..5764 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5680..5699) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5734..5753 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5813..5829) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5813..5829) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5826..5848) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5837..5853 /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(5861..5891) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5906..5927 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6044..6061) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6215..6800) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6295..6314) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6971..7831) /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 7594..7613 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7832..7936) /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 atatggatcc 1081 tctgaatctg tcatggtatg acgatgatct ggaaaggcaa aactggtccc ggccgtttaa 1141 tggatcagat ggtaaagcag atcggcctca ctacaattac tacgccacac tgctcaccct 1201 cctcattgca gttattgtgt tcgggaacgt gctggtttgc atggcagtaa gccgggaaaa 1261 ggctctccag acaactacca actacctcat cgtctcactt gcggtagcag acttgctggt 1321 ggctaccctg gtcatgcctt gggtggttta cttggaggtt gttggcgagt ggaagttctc 1381 aaggatacat tgcgatatct ttgtaaccct ggatgttatg atgtgtacgg cttcaatatt 1441 gaacctctgt gccatctcca tcgaccgcta tactgccgta gcaatgccta tgctttacaa 1501 tacccgatat tcaagcaagc ggagggtgac ggttatgatt tccatcgttt gggtgctgtc 1561 ctttacgatt agttgccctc tgctgttcgg gctgaacaac gcagaccaaa atgagtgtat 1621 tattgccaat cccgcctttg tggtgtattc atcaattgtg agcttttacg tcccatttat 1681 tgttacattg ttggtgtaca ttaaaatata cattgtcctc aggcggcgga gaaaacgggt 1741 gaacaccaag cggtcatcta gagctttccg cgctcatctg agagctccac tgaagggcaa 1801 ctgtacccac ccggaagaca tgaaactgtg taccgtcatc atgaaatcca atgggagctt 1861 ccccgttaat aggcggagag tggaagccgc aaggcgcgca caggagctgg agatggagat 1921 gctcagcagc accagccccc ctgagcgcac ccggtacagc cccattccgc cctcacatca 1981 ccaactcaca ctgcccgacc cctcacatca tggactccac tctaccccgg actcaccggc 2041 caagcctgaa aagaacgggc acgcgaaaga tcacccgaaa atcgctaaga ttttcgagat 2101 ccaaactatg cctaacggca aaactcgcac ctctctgaaa acaatgagtc ggagaaaact 2161 ttcacagcaa aaggagaaga aggccaccca aatgctggcc attgtactcg gggtgttcat 2221 catttgttgg ctgccctttt ttatcaccca catcctgaac attcattgtg attgcaacat 2281 tccaccagtg ctctatagcg cctttacttg gctgggttat gtgaatagtg cggtaaaccc 2341 cataatctac accacattca atatcgaatt tcgaaaagca tttctgaaga ttctccactg 2401 tatcgatacc ggtggacgca ccccacccag cctgggtccc caagatgagt cctgcaccac 2461 cgccagctcc tccctggcca aggacacttc atcgaccggt gagaacctgt acttccagct 2521 aagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat 2581 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta 2641 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag 2701 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa 2761 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg 2821 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca 2881 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt 2941 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac 3001 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc 3061 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa 3121 atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa aacaattacg ggtctaccat 3181 cgagggcctg ctcgatctcc cggacgacga cgcccccgaa gaggcggggc tggcggctcc 3241 gcgcctgtcc tttctccccg cgggacacac gcgcagactg tcgacggccc ccccgaccga 3301 tgtcagcctg ggggacgagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga 3361 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt 3421 taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca 3481 gatgtttacc gatgcccttg gaattgacga gtacggtggg tagctcgagt ctagagggcc 3541 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 3601 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 3661 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 3721 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt 3781 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc 3841 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3901 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 3961 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 4021 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 4081 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 4141 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 4201 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 4261 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta 4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga 4621 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt 4681 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 4741 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4801 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac 4861 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4921 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4981 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 5041 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 5101 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 5161 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag 5221 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 5281 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 5341 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 5401 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 5461 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 5521 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 5581 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 5641 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 5701 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 5761 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 5821 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 5881 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 5941 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 6001 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 6061 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 6121 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 6181 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 6241 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 6301 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 6361 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 6421 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 6481 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 6541 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 6601 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 6661 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 6721 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc 6781 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 6841 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 6901 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 6961 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 7021 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 7081 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 7141 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 7201 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 7261 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 7321 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 7381 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 7441 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 7501 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 7561 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 7621 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 7681 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 7741 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 7801 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 7861 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 7921 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //