LOCUS       sequence-209676-        7621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a chimeric G-protein coupled receptor 
            (Rhodopsin-GPR149; Opto-GPR149; E3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7621)
  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 7621)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-05-16"; volume: "9(1):1950. doi"; pages: " 
            10.1038/s41467-018-04342-1"
COMMENT     Sequence Label: Opto-GPR149
FEATURES             Location/Qualifiers
     source          1..7621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(3051..3068)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3057..3281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3327..3755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3414..3433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3624..3645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3764..3784)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3769..4098
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3949..4084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4011..4030
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4165..4959
                     /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(4219..4238)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4829..4848
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5133..5254
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5170..5189)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5224..5243
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5303..5319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5303..5319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5316..5338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5327..5343
                     /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(join(5351..5357,5358..5375,5376..5381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5396..5417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5534..5551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5705..6293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5785..5804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6464..7255,7256..7324))
                     /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     7087..7106
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7325..7429)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7504..7523)
                     /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 acgtcctgct gaaaatgcag aacagaactg ttctcaacta
     1081 catcctgctc aacctggccg tggccgacct cttcatggtg ttcgggggct tcaccaccac
     1141 cctctacacc tctctgcacg ggtacttcgt ctttgggccc acgggctgca acctggaggg
     1201 cttctttgcc accttgggcg gtgaaattgc actgtggtcc ttggtggtcc tggccatcga
     1261 gcggtacgtg gtggtgcaca gaggtgtggg gagccagaca gcctccagaa gatcgggcgc
     1321 catcatgggc gtcgccttca cctgggtcat ggctctggcc tgtgccgcgc cccccctcgt
     1381 cggctggtcc aggtacatcc cggagggcat gcagtgctcg tgcgggattg actactacac
     1441 gccccacgag gaaaccaaca atgagtcgtt cgtcatctac atgttcgtgg tccacttcat
     1501 catccccctg attgtcatat tcttctgcta cggagaaatt tcccgtggag cttcaattcc
     1561 tgggacccct cctactgcgg ggagagtggt ttccctgtcc ccagaggatg ctccaggccc
     1621 gagtctgcgg cgctctgggg gatgctctcc gagctccgac accgtgttcg gaccgggtgc
     1681 gcccgctgcc gctggggctg aagcctgcag gcgtgagaac cgggggactc tctatggcac
     1741 caggagcttc accgtgagcg tagcgcagaa gcgcatggtg atcatcatgg tcatcgcttt
     1801 cctaatctgc tggctgccct acgctggggt ggcgttctac atcttcaccc atcagggctc
     1861 tgactttggc cccatcttca tgaccatccc ggctttcttt gccaagactt ctgccgtcta
     1921 caaccccgtc atctacatca tgatgaacaa gcagttccgc ctatcattcc aaaaaagtta
     1981 tgggatttat aaaatagcac atgaagatta ctatgatgat gatgaaaatt ccatattcta
     2041 tcacaacctg atgaactctg agtgtgaaac tacaaaagac cctcagagag acaaccgtaa
     2101 catcttcaat gctataaaag tagaaatcag caccacgccc tctctggaca gctccacaca
     2161 aagaggcatc aacaaatgca caaatactga tattacagaa gctaaacagg attccaacaa
     2221 caaaaaggat gcgttttctg acaaaacagg aggtgatatt aactatgaag aaactacctt
     2281 ttctgaaggg ccagaaagaa ggctgtctca tgaagagagt cagaaaccag atctttcaga
     2341 ctgggagtgg tgtaggagta aatcagaaag aacccctcgt cagcgttccg gttatgccct
     2401 tgccattccc ttgtgtgcat tccaggggac tgtgtctctc catgcaccta cagggaaaac
     2461 cctatctctt tctacctatg aggtaagcgc agaagggcaa aaaataactc cagcctctaa
     2521 gaaaatagaa gtctatcgat ccaaaagtgt tggccatgaa ccaaactcag aagattcttc
     2581 atccacgttt gtggacacca gtgtgaaaat acacttggag gttcttgaaa tttgtgataa
     2641 tgaagaggcc ttggacactg tgtcaatcat tagtaacatc agtcagtcct ccacacaagt
     2701 cagatctcca tccctacgtt actccaggaa agaaaacaga tttgtttcat gtgacctagg
     2761 ggaaacagcc tcatactccc tctttttgcc caccagtaat cctgatggtg atattaatat
     2821 ctccattcca gacacagtag aagcacacag gcagaacagt aaaaggcagc atcaagagag
     2881 ggatggctac caggaggaaa tccagttgtt aaataaagct tacagaaaaa gagaggaaga
     2941 aagcaagggt agtacagaaa ccagccaagt ggcgcctgcc taagaattcc accacactgg
     3001 actagtggat ccgagctcgg taccaagctt aagtttaaac cgctgatcag cctcgactgt
     3061 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3121 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3181 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3241 agagaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3301 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3361 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3421 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3481 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3541 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3601 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3661 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3721 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3781 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3841 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3901 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3961 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4021 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4081 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4141 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4201 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4261 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4321 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4381 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4441 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4501 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4561 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4621 gtcgatcagg atgatctgga cgaagaacat caggggctcg cgccagccga actgttcgcc
     4681 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4741 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4801 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagaactt
     4861 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4921 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4981 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5041 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5101 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5161 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5221 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5281 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5341 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5401 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5461 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5521 gctcttgcgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5581 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5641 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5701 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5761 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5821 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5881 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5941 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6001 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6061 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6121 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6181 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6241 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6301 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6361 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6421 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6481 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6541 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6601 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6661 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6721 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6781 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6841 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6901 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6961 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7021 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7081 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7141 cttcggggcg aaaactctca aggatcttac cactgttgag atccagttcg atgtaaccca
     7201 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7261 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7321 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7381 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7441 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     7501 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7561 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7621 c
//
