LOCUS       Exported                5091 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR150; Opto-GPR150; E4).
ACCESSION   .
VERSION     .
KEYWORDS    Opto-GPR150
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5091)
  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 5091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5091
                     /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(2157..2174)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2163..2387
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2603..2724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2640..2659)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2694..2713
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2773..2789)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2773..2789)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2786..2808)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2797..2813
                     /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(2821..2851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2866..2887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3004..3021)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3175..3763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3255..3274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3934..4794)
                     /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     4557..4576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4795..4899)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4974..4993)
                     /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 acgtcctgtg cggcggcggc gggccctggg cgggccccaa
     1081 gcgtcgcaag ctcaactaca tcctgctcaa cctggccgtg gccgacctct tcatggtgtt
     1141 cgggggcttc accaccaccc tctacacctc tctgcacggg tacttcgtct ttgggcccac
     1201 gggctgcaac ctggagggct tctttgccac cttgggcggt gaaattgcac tgtggtcctt
     1261 ggtggtcctg gccatcgagc ggtacgtggt ggtgcggctc ccgcacggcc ggccgctgcc
     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 ctacggacac ctactctccg tctggtggcg
     1561 gcaccggccg caggcccccg cggctgcagc gccctggtcg gcgagcccag gtcgagcccc
     1621 tgcgcccagc gcgctgcccc gcgccaaggt gcagagccgc atggtgatca tcatggtcat
     1681 cgctttccta atctgctggc tgccctacgc tggggtggcg ttctacatct tcacccatca
     1741 gggctctgac tttggcccca tcttcatgac catcccggct ttctttgcca agacttctgc
     1801 cgtctacaac cccgtcatct acatcatgat gaacaagcag ttccgccgac agctgcggaa
     1861 gcggctgggc tctctgtgct gcgcgccgca gggaggcgcg gaggacgagg aggggccccg
     1921 gggccaccag gcgctctacc gccaacgctg gccccaccct cattatcacc atgctcggcg
     1981 ggaaccgctg gacgagggcg gcttgcgccc accccctccg cgccccagac ccctgccttg
     2041 ctcctgcgaa agtgccttca cagaaaccag ccaagtggcg cctgcctaag aattccacca
     2101 cactggacta gtggatccga gctcggtacc aagcttaagt ttaaaccgct gatcagcctc
     2161 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2221 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2281 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2341 ttgggaagag aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2401 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2461 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgccttct atgaaaggtt
     2521 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     2581 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     2641 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2701 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     2761 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2821 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2881 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2941 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttgcgc
     3001 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3061 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3121 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3181 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3241 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3301 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3361 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3421 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3481 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3541 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3601 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3661 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3721 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3781 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3841 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3901 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3961 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4021 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc
     4081 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4141 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4201 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4261 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4321 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4381 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4441 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4501 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4561 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4621 aaaactctca aggatcttac cactgttgag atccagttcg atgtaaccca ctcgtgcacc
     4681 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4741 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4801 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4921 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     4981 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     5041 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga c
//
