LOCUS Exported 8066 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 DRD5-Tango SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8066) 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 8066) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8066 /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 2390..3391 /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(2457..2476) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(3423..3440) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3429..3653 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3699..4127 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3786..3805) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3996..4017 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4136..4156) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4141..4470 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4321..4456 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4383..4402 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4537..5331 /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(4591..4610) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5201..5220 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5505..5626 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5542..5561) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5596..5615 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5675..5691) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5675..5691) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5688..5710) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5699..5715 /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(5723..5753) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5768..5789 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5906..5923) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6077..6665) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6157..6176) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6836..7696) /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 7459..7478 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7697..7801) /gene="bla" /label=AmpR promoter primer_bind complement(7876..7895) /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 gttgccacca gggagcaatg gtacggcgta tcccgggcag ttcgccctct accaacaact 901 tgcccagggt aatgccgtcg ggggcagcgc aggcgctcca cctttggggc caagtcaggt 961 cgtgactgcc tgtcttctga cccttctgat aatatggact ctgctgggca acgtcctggt 1021 ttgtgcagca attgtgcgct ctcgccacct gcgagctaac atgacaaacg tgttcatagt 1081 gagcctggca gtgagcgacc tgttcgttgc gctcctcgtg atgccctgga aggcggtggc 1141 agaagtggcc ggatattggc cttttggcgc attttgcgat gtttgggtgg cctttgatat 1201 tatgtgttcc acggccagta ttctcaatct gtgcgtgatt tccgtcgatc gctactgggc 1261 aattagcagg ccgttccgct ataaaagaaa gatgacccag aggatggccc ttgtgatggt 1321 cggactggcc tggaccctct ccatcctgat ttctttcatt cctgttcagt tgaattggca 1381 cagggaccag gctgcaagtt ggggcgggct ggatctgcca aataacctgg ccaattggac 1441 cccatgggag gaagatttct gggaaccaga cgtcaacgca gagaattgtg atagtagcct 1501 taatcggacc tatgctatta gttccagtct gatctccttc tacattccgg tcgctataat 1561 gatagtgact tacacaagga tctatagaat cgctcaagtt caaatacgaa ggatctctag 1621 cctggagcgc gccgctgagc acgctcaatc ctgccgctca tccgctgctt gtgctcccga 1681 cacctctctc agagcttcta tcaagaaaga aaccaaggtc ctcaagactc tgagcgtgat 1741 tatgggcgtc ttcgtgtgct gttggctccc cttcttcatt ctgaattgca tggtcccatt 1801 ttgctctggg cacccagaag ggccccccgc agggtttcct tgtgtaagtg agacgacttt 1861 cgacgtcttt gtgtggtttg ggtgggcaaa tagctctttg aatcctgtaa tctacgcatt 1921 taacgctgat tttcagaaag tcttcgcgca gctgctgggt tgtagtcatt tttgctcacg 1981 aacgcctgtg gaaacagtca acatctctaa tgaactcatc tcctacaacc aggacatcgt 2041 attccacaaa gagattgcag ctgcatacat ccacatgatg cccaatgctg ttacacctgg 2101 aaacagggag gtggataacg acgaggagga gggccccttc gatcggatgt tccagattta 2161 tcagacgagc cctgacggcg acccagtcgc tgaatctgtg tgggagctgg actgtgaagg 2221 tgaaatatca ctggataaaa tcactccctt tacgcccaac ggatttcaca tcgataccgg 2281 tggacgcacc ccacccagcc tgggtcccca agatgagtcc tgcaccaccg ccagctcctc 2341 cctggccaag gacacttcat cgaccggtga gaacctgtac ttccagctaa gattagataa 2401 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac 2461 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa 2521 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca 2581 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt 2641 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac 2701 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc 2761 actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt 2821 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat 2881 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt 2941 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg 3001 gtccgcgtac agccgcgcgc gtacgaaaaa caattacggg tctaccatcg agggcctgct 3061 cgatctcccg gacgacgacg cccccgaaga ggcggggctg gcggctccgc gcctgtcctt 3121 tctccccgcg ggacacacgc gcagactgtc gacggccccc ccgaccgatg tcagcctggg 3181 ggacgagctc cacttagacg gcgaggacgt ggcgatggcg catgccgacg cgctagacga 3241 tttcgatctg gacatgttgg gggacgggga ttccccgggt ccgggattta ccccccacga 3301 ctccgccccc tacggcgctc tggatatggc cgacttcgag tttgagcaga tgtttaccga 3361 tgcccttgga attgacgagt acggtgggta gtctagaggg cccgtttaaa cccgctgatc 3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 3661 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 3841 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 3901 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 3961 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 4021 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 4081 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 4381 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 4441 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc 4501 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac 4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca 4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt 4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg 4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga 4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct 4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg 4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg 4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc 5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat 5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac 5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt 5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct 5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc 5341 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca 5401 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga 5461 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag 5521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt 5581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac 5641 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt 5701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg 5761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt 5821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt 5881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc 5941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg 6001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg 6061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac 6121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg 6181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct 6241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg 6301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct 6361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac 6421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt 6481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc 6541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca 6601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat 6661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac 6721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt 6781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc 6841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg 6901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg 6961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc 7021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta 7081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg 7141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct 7201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta 7261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg 7321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga 7381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt 7441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca 7501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt 7561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt 7621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga 7681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt 7741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc 7801 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct 7861 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc 7921 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca 7981 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg 8041 cgatgtacgg gccagatata cgcgtt //