LOCUS       sequence-387100-        7763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse SMO with an N-terminal HiBiT; contains D54N and 
            R690G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7763)
  AUTHORS   Turku A, Schihada H, Kozielewicz P, Bowin CF, Schulte G
  TITLE     Residue 6.43 defines receptor function in class F GPCRs.
  JOURNAL   Nat Commun. 2021 Jun 24;12(1):3919. doi: 10.1038/s41467-021-24004-z.
  PUBMED    34168128
REFERENCE   2  (bases 1 to 7763)
  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: "2021-06-24"; volume: "12(1):3919. doi"; pages: " 
            10.1038/s41467-021-24004-z"
COMMENT     Sequence Label: HiBiT-SMO
FEATURES             Location/Qualifiers
     source          1..7763
                     /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"
     primer_bind     complement(3354..3371)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3360..3584
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3630..4058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3717..3736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3927..3948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4067..4087)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4072..4401
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4252..4387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4314..4333
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4468..5262
                     /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(4522..4541)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5132..5151
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5436..5557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5473..5492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5527..5546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5606..5622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5606..5622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5619..5641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5630..5646
                     /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(5654..5660,5661..5678,5679..5684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5837..5854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6008..6596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6088..6107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6767..7558,7559..7627))
                     /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     7390..7409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7628..7732)
                     /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 gtttaaactt aagcttgcca ccatgcggct ctgcatcccg caggtgctgt tggccttgtt
      961 cctttccatg ctgacagggc cgggagaagg cagccgggtg agcggctggc ggctgttcaa
     1021 gaagattagc ggatccgcgg ccttgagcgg gaacgtgacc gggcctgggc ctcacagcgc
     1081 cagcgggagc tcgaggagga acgtgccggt gaccagccct ccgccgccgc tgctgagcca
     1141 ctgcggccgg gccgcccact gcgagccttt gcgctacaac gtgtgcctgg gctcggcgct
     1201 gccctacgga gccaccacca cgctgctggc tggggactcg gactcgcagg aggaagcgca
     1261 cggcaagctc gtgctctggt ccggcctccg gaatgccccc cgctgctggg cagtgatcca
     1321 gcccctgctg tgtgctgtct acatgcccaa gtgtgagaat gaccgagtgg agttgcccag
     1381 ccgtaccctc tgccaggcca cccgaggccc ctgtgccatt gtggagcggg agcgagggtg
     1441 gcctgacttt ctgcgttgca caccggacca cttccctgaa ggctgcccaa acgaggtaca
     1501 aaacatcaag ttcaacagct caggccaatg tgaagcaccc ttggtgcgaa cagacaaccc
     1561 caagagctgg tatgaggacg tggagggctg tgggattcag tgtcagaacc cgctgttcac
     1621 cgaggccgag caccaggaca tgcacagcta catcgcagcc ttcggggcgg tcactggtct
     1681 ctgcacgctc ttcacactgg ccacctttgt ggctgactgg cggaattcca atcgctaccc
     1741 tgcggttatt ctcttctatg tcaatgcgtg tttcttcgtg ggcagcattg gctggctggc
     1801 ccagttcatg gatggtgccc gccgagagat tgtttgccga gcagatggca ccatgagatt
     1861 tggggagccc acctccagtg agaccctgtc ctgtgtcatc atctttgtca ttgtgtacta
     1921 tgccttgatg gctggagtag tctggttcgt ggtcctcacc tatgcctggc acacctcctt
     1981 caaagccctg ggcaccacct accagcctct ctcgggcaag acatcctatt tccacctgct
     2041 cacgtggtca ctcccctttg tcctcacggt ggcaatcctg gctgtggctc aggtagatgg
     2101 agactccgtg agtggcatct gttttgtagg ctacaagaac tatcggtacc gtgctggctt
     2161 tgtcctggcc ccaattggcc tggtgcttat tgtgggaggc tacttcctca tcagaggggt
     2221 catgactctg ttctccatca agagcaacca ccctgggctc ttaagtgaga aggcagccag
     2281 caagatcaac gagaccatgc tgcgcctggg catttttggc ttcctggcct ttggctttgt
     2341 gctcatcacc ttcagctgcc acttctatga cttcttcaac caggctgagt gggagcgtag
     2401 cttccgggac tatgtgctat gccaagccaa cgtgaccatc gggctgccta ccaagaagcc
     2461 cattcctgac tgtgagatca agaatcggcc cagcctcctg gtggagaaga tcaatctatt
     2521 tgccatgttt ggcactggca ttgccatgag cacctgggtc tggaccaagg ccaccctgct
     2581 catctggagg cgcacctggt gcaggttgac tgggcacagt gatgatgagc ccaagagaat
     2641 caagaagagc aagatgatcg ccaaggcctt ctctaagcgg cgtgagctgc tgcagaaccc
     2701 gggccaggag ctctccttca gcatgcacac tgtctcccat gatggacctg ttgcgggttt
     2761 ggcttttgac ctcaatgaac cctcagctga cgtctcctct gcctgggctc agcatgtcac
     2821 caagatggtg gctcggagag gagccatatt gccccaggat gtgtccgtta cccctgtggc
     2881 aactccagtg ccaccagaag aacaagccaa catgtggctg gttgaggcag agatctcccc
     2941 agagttagag aagcgtttgg gccggaagaa aaagcggagg aagaggaaga aggaggtgtg
     3001 ccccttgggg ccagcccctg agcttcacca ctctgcccct gttcctgcca ccagtgcagt
     3061 tcctcggctg cctcagctgc ctcggcagaa gtgcctggta gctgcaaacg cctggggaac
     3121 aggggagtcc tgccgacagg gagcctggac tctagtctcc aaccccttct gcccagagcc
     3181 tagtccccat caagatccat ttctccctgg tgcctcagcc ccccgggtct gggctcaggg
     3241 ccgcctccag gggctggggt ccattcattc ccgcactaac ctaatggagg ctgagatctt
     3301 ggatgcagac tcggacttct gatctagagg gcccgtttaa acccgctgat cagcctcgac
     3361 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3421 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3481 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3541 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3601 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3661 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3721 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3781 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3841 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3901 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3961 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4021 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4081 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4141 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4201 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4261 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4321 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4381 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4441 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4501 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4561 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4621 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4681 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4741 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4801 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4861 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4921 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4981 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5041 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5101 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5161 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5221 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5281 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5341 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5401 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5461 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5521 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5581 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5641 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5701 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5761 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5821 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5881 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5941 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6421 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6541 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6661 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6721 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7741 cccgaaaagt gccacctgac gtc
//
