LOCUS Exported 6568 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of a chimeric G-protein coupled receptor (Rhodopsin-GPR37; Opto-GPR37; B9). ACCESSION . VERSION . KEYWORDS Opto-GPR37 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6568) AUTHORS Morri M, Sanchez-Romero I, Tichy AM, Kainrath S, Gerrard EJ, Hirschfeld PP, Schwarz J, Janovjak H TITLE Optical functionalization of human Class A orphan G-protein-coupled receptors. JOURNAL Nat Commun. 2018 May 16;9(1):1950. doi: 10.1038/s41467-018-04342-1. PUBMED 29769519 REFERENCE 2 (bases 1 to 6568) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6568 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 74..453 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 454..657 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 608..628 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 702..721 /label=T7 /note="T7 promoter, forward primer" promoter 702..720 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 821..853 /codon_start=1 /product="epitope tag from vesicular stomatitis virus G protein" /label=VSV-G tag /translation="YTDIEMNRLGK" CDS 878..916 /codon_start=1 /product="segment of bovine rhodopsin containing a signal for N-linked glycosylation (Favaloro et al., 2008)" /label=opsin tag /note="The asparagine in the NKT tripeptide will be glycosylated if this sequence is exposed to the ER lumen." /translation="GPNFYVPFSNKTG" primer_bind complement(2076..2093) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2082..2306 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2352..2780 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2439..2458) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2649..2670 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2789..2809) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2794..3123 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2974..3109 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3036..3055 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3244..3263) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3776..3795 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4080..4201 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4117..4136) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4171..4190 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4250..4266) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4250..4266) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4263..4285) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4274..4290 /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(4298..4328) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4343..4364 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4481..4498) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4652..5240) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4732..4751) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5411..6271) /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 6034..6053 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6272..6376) /gene="bla" /label=AmpR promoter primer_bind complement(6451..6470) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc 61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca 121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc 661 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg 721 gagacacaag ctggctagcg tttaaacggg ccctctagac tcgagcacca tgaaaacgat 781 catcgccctg agctacatct tctgcctggt attcgccatg tacaccgata tagagatgaa 841 caggctggga aaggatagcc tcatgaacgg gaccgagggc ccaaacttct acgtgccttt 901 ctccaacaag acgggcgtgg tgcgcagccc cttcgaggcc ccgcagtact acctggcgga 961 gccatggcag ttctccatgc tggccgccta catgttcctg ctgatcatgc ttggcttccc 1021 catcaacttc ctcacgctgt acgtcgtgtg ccacaactac tacatgcgga gcatcctcaa 1081 ctacatcctg ctcaacctgg ccgtggccga cctcttcatg gtgttcgggg gcttcaccac 1141 caccctctac acctctctgc acgggtactt cgtctttggg cccacgggct gcaacctgga 1201 gggcttcttt gccaccttgg gcggtgaaat tgcactgtgg tccttggtgg tcctggccat 1261 cgagcggtac gtggtggtga ccaacgtaca gatgtactac gaaatgatcg aaaactgttc 1321 cgccatcatg ggcgtcgcct tcacctgggt catggctctg gcctgtgccg cgccccccct 1381 cgtcggctgg tccaggtaca tcccggaggg catgcagtgc tcgtgcggga ttgactacta 1441 cacgccccac gaggaaacca acaatgagtc gttcgtcatc tacatgttcg tggtccactt 1501 catcatcccc ctgattgtca tattcttctg ctacggagtg actgcgagga aaatccgcaa 1561 agcagagaaa gcctgtaccc gagggaataa acggcagatt caactagaga gtcagatgaa 1621 ccgcatggtg atcatcatgg tcatcgcttt cctaatctgc tggctgccct acgctggggt 1681 ggcgttctac atcttcaccc atcagggctc tgactttggc cccatcttca tgaccatccc 1741 ggctttcttt gccaagactt ctgccgtcta caaccccgtc atctacatca tgatgaacaa 1801 gcagttccgc cgggccttca tggagtgctg ctgctgttgc tgtgaggaat gcattcagaa 1861 gtcctcaacg gtgaccagtg atgacaatga caacgagtac accacggaac tcgaactctc 1921 gcctttcagt accatacgcc gtgaaatgtc cacttttgct tctgtcggaa ctcattgcac 1981 agaaaccagc caagtggcgc ctgcctaaga attccaccac actggactag tggatccgag 2041 ctcggtacca agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca 2101 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 2161 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 2221 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca 2281 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 2341 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 2401 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 2461 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 2521 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 2581 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 2641 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 2701 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 2761 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 2821 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 2881 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2941 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 3001 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 3061 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 3121 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 3181 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 3241 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 3301 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 3361 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 3421 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 3481 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 3541 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 3601 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 3661 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3721 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3781 gctacccgtg atattgctga agaacttggc ggcgaatggg ctgaccgctt cctcgtgctt 3841 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3901 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3961 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4021 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4201 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4441 cggggagagg cggtttgcgt attgggcgct cttgcgcttc ctcgctcact gactcgctgc 4501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5521 catctggccc cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat 5581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5701 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6001 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccac 6121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6361 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 6421 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag 6481 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta 6541 agctacaaca aggcaaggct tgaccgac //