LOCUS       sequence-494487-        6729 bp    DNA     circular SYN 03-DEC-2025
DEFINITION  Express GPR6 in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6729)
  AUTHORS   Barekatain M, Johansson LC, Lam JH, Chang H, Sadybekov AV, Han GW, 
            Russo J, Bliesath J, Brice NL, Carlton MBL, Saikatendu KS, Sun H, 
            Murphy ST, Monenschein H, Schiffer HH, Popov P, Lutomski CA, 
            Robinson CV, Liu ZJ, Hua T, Katritch V, Cherezov V
  TITLE     Structural insights into the high basal activity and inverse agonism
            of the orphan receptor GPR6 implicated in Parkinson's disease.
  JOURNAL   Sci Signal. 2024 Dec 3;17(865):eado8741. doi: 
            10.1126/scisignal.ado8741. Epub 2024 Dec 3.
  PUBMED    39626010
REFERENCE   2  (bases 1 to 6729)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Signal."; date: "2024-12-3"; volume: "17(865):eado8741. doi"; pages:
            " 10.1126/scisignal.ado8741. Epub 2024 Dec 3"
COMMENT     Sequence Label: IMPT-10917
FEATURES             Location/Qualifiers
     source          1..6729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             2264..2290
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(2320..2337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2326..2550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2596..3024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2683..2702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2893..2914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3033..3053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3038..3367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3218..3353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3280..3299
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3434..4228
                     /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(3488..3507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4098..4117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4402..4523
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4439..4458)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4493..4512
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4572..4588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4572..4588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4585..4607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4596..4612
                     /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(4620..4626,4627..4644,4645..4650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4665..4686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4803..4820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4974..5562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5054..5073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5733..6524,6525..6593))
                     /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     6356..6375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6594..6698)
                     /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 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcc
      961 accacactgg actagtggat ccatgaaaac tattattgcc ctgagctaca tcttctgcct
     1021 ggtctttgcc ggcggcgcca atggcagcct ggagctgagc tcccagctgt ccgccggacc
     1081 accaggcctg ctgctgccag ccgtgaaccc ttgggacgtg ctgctgtgcg tgagcggaac
     1141 cgtgatcgca ggagagaatg cactggtggt ggccctgatc gcaagcaccc cagccctgag
     1201 gacacccatg ttcgtgctgg tgggctccct ggcaacagca gatctgctgg caggactcgg
     1261 cctgatcctg cacttcgtgt ttcagtacct ggtgcccagc gagacagtga gcctgctgac
     1321 agtgggcttc ctggtggcct cttttgccgc cagcgtgtct agcctgctgg ccatcaccgt
     1381 ggaccggtac ctgtctctgt ataaccccct gacctactat agccggagaa cactgctggg
     1441 agtgcacctg ctgctggcag caacctggac agtgtctctg ggcctgggcc tgctgccagt
     1501 gctgggatgg aattgcctgg cagagagagc agcatgttcc gtggtgaggc ccctggcacg
     1561 ctcccacgtg gccctgctgt ctgccgcctt ctttatggtg ttcggcatca tgctgcacct
     1621 gtacgtgagg atctgccagg tggtgtggag gcacgcagcc gacctggaag ataactggga
     1681 gacactgaac gacaacctga aggtgatcga gaaggccgat aatgccgctc aagtgaagga
     1741 cgccctgacc aagatgcggg ccgctgctct cgatgctcag aaagccaccc ctcctaagct
     1801 ggaagataag tctccagaca gccccgagat gaaagacttc agacacggct tcgacatcct
     1861 ggtgggccag atcgacgacg ccctgaaact ggccaacgag ggaaaggtca aggaagccca
     1921 ggccgccgcc gagcagctga agaccacaag aaacgcctac atccagaagt acctggccca
     1981 cctggcagca accaggaagg gagtgcggac actggcagtg gtgctgggca ccttcggcgc
     2041 atgctggctg ccctttgcca tctattgcgt ggtgggctct cacgaggacc ccgccgtgta
     2101 cacctatgcc acactgctgc ctgccacact gaactccatg atcaatccaa tcatctatgc
     2161 ctttcggaac caggagatcc agagagccct gtggctgctg ctggacgggt gtttccagag
     2221 caaggtgcca ttcaggagca ggtccccaag cgaagtcgag ttccaccacc accaccacca
     2281 ccaccaccac taaaagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt
     2341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2521 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     2581 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3001 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3121 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3181 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3241 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3301 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3361 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     3421 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3481 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3541 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3601 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3661 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3721 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3781 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3841 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3901 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3961 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4021 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4081 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4141 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4201 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4261 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4321 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4381 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     4441 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4501 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4561 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4621 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4681 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4741 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5401 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5581 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5641 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5701 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5761 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5821 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5881 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5941 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6001 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6061 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6121 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6181 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6241 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6301 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6361 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6421 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6481 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6541 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6601 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6661 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6721 cctgacgtc
//
