LOCUS       sequence-233148-        7733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses wildtype Ptch1 for function assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7733)
  AUTHORS   Zhang Y, Bulkley DP, Xin Y, Roberts KJ, Asarnow DE, Sharma A, Myers 
            BR, Cho W, Cheng Y, Beachy PA
  TITLE     Structural Basis for Cholesterol Transport-like Activity of the 
            Hedgehog Receptor Patched.
  JOURNAL   Cell. 2018 Nov 15;175(5):1352-1364.e14. doi: 
            10.1016/j.cell.2018.10.026. Epub 2018 Nov 8.
  PUBMED    30415841
REFERENCE   2  (bases 1 to 7733)
  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"; doi: 
            "10.1016/j.cell.2018.10.026"; journalName: "Cell"; date: 
            "2018-11-15- 15"; volume: "175"; issue: "5"; pages: "1352-1364.e14"
COMMENT     Sequence Label: pcDNA-h-mmPtch1-FL
FEATURES             Location/Qualifiers
     source          1..7733
                     /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"
     promoter        complement(5224..5242)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5225..5242)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5262..5279)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5268..5492
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5576..5592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5576..5592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5589..5611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5600..5616
                     /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(5624..5630,5631..5648,5649..5654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5669..5690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5807..5824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6737..7528,7529..7597))
                     /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     7360..7379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7598..7702)
                     /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 gtactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 accatggcct cggctggtaa cgccgccggg gccctgggca ggcaggccgg cggcgggagg
      961 cgcagacgga ccgggggacc gcaccgcgcc gcgccggacc gggactatct gcaccggccc
     1021 agctactgcg acgccgcctt cgctctggag cagatttcca aggggaaggc tactggccgg
     1081 aaagcgccgc tgtggctgag agcgaagttt cagagactct tatttaaact gggttgttac
     1141 attcaaaaga actgcggcaa gtttttggtt gtgggtctcc tcatatttgg ggccttcgct
     1201 gtgggattaa aggcagctaa tctcgagacc aacgtggagg agctgtgggt ggaagttggt
     1261 ggacgagtga gtcgagaatt aaattatacc cgtcagaaga taggagaaga ggctatgttt
     1321 aatcctcaac tcatgataca gactccaaaa gaagaaggcg ctaatgttct gaccacagag
     1381 gctctcctgc aacacctgga ctcagcactc caggccagtc gtgtgcacgt ctacatgtat
     1441 aacaggcaat ggaagttgga acatttgtgc tacaaatcag gggaacttat cacggagaca
     1501 ggttacatgg atcagataat agaatacctt tacccttgct taatcattac acctttggac
     1561 tgcttctggg aaggggcaaa gctacagtcc gggacagcat acctcctagg taagcctcct
     1621 ttacggtgga caaactttga ccccttggaa ttcctagaag agttaaagaa aataaactac
     1681 caagtggaca gctgggagga aatgctgaat aaagccgaag ttggccatgg gtacatggac
     1741 cggccttgcc tcaacccagc cgacccagat tgccctgcca cagcccctaa caaaaattca
     1801 accaaacctc ttgatgtggc ccttgttttg aatggtggat gtcaaggttt atccaggaag
     1861 tatatgcatt ggcaggagga gttgattgtg ggtggtaccg tcaagaatgc cactggaaaa
     1921 cttgtcagcg ctcacgccct gcaaaccatg ttccagttaa tgactcccaa gcaaatgtat
     1981 gaacacttca ggggctacga ctatgtctct cacatcaact ggaatgaaga cagggcagcc
     2041 gccatcctgg aggcctggca gaggacttac gtggaggtgg ttcatcaaag tgtcgcccca
     2101 aactccactc aaaaggtgct tcccttcaca accacgaccc tggacgacat cctaaaatcc
     2161 ttctctgatg tcagtgtcat ccgagtggcc agcggctacc tactgatgct tgcctatgcc
     2221 tgtttaacca tgctgcgctg ggactgctcc aagtcccagg gtgccgtggg gctggctggc
     2281 gtcctgttgg ttgcgctgtc agtggctgca ggattgggcc tctgctcctt gattggcatt
     2341 tcttttaatg ctgcgacaac tcaggttttg ccgtttcttg ctcttggtgt tggtgtggat
     2401 gatgtcttcc tcctggccca tgcattcagt gaaacaggac agaataagag gattccattt
     2461 gaggacagga ctggggagtg cctcaagcgc accggagcca gcgtggccct cacctccatc
     2521 agcaatgtca ccgccttctt catggccgca ttgatcccta tccctgccct gcgagcgttc
     2581 tccctccagg ctgctgtggt ggtggtattc aattttgcta tggttctgct catttttcct
     2641 gcaattctca gcatggattt atacagacgt gaggacagaa gattggatat tttctgctgt
     2701 ttcacaagcc cctgtgtcag cagggtgatt caagttgagc cacaggccta cacagagcct
     2761 cacagtaaca cccggtacag ccccccaccc ccatacacca gccacagctt cgcccacgaa
     2821 acccatatca ctatgcagtc caccgttcag ctccgcacag agtatgaccc tcacacgcac
     2881 gtgtactaca ccaccgccga gccacgctct gagatctctg tacagcctgt taccgtcacc
     2941 caggacaacc tcagctgtca gagtcccgag agcaccagct ctaccaggga cctgctctcc
     3001 cagttctcag actccagcct ccactgcctc gagcccccct gcaccaagtg gacactctct
     3061 tcgtttgcag agaagcacta tgctcctttc ctcctgaaac ccaaagccaa ggttgtggta
     3121 atccttcttt tcctgggctt gctgggggtc agcctttatg ggaccacccg agtgagagac
     3181 gggctggacc tcacggacat tgttccccgg gaaaccagag aatatgactt catagctgcc
     3241 cagttcaagt acttctcttt ctacaacatg tatatagtca cccagaaagc agactacccg
     3301 aatatccagc acctacttta cgaccttcat aagagtttca gcaatgtgaa gtatgtcatg
     3361 ctggaggaga acaagcaact tccccaaatg tggctgcact actttagaga ctggcttcaa
     3421 ggacttcagg atgcatttga cagtgactgg gaaactggga ggatcatgcc aaacaattat
     3481 aaaaatggat cagatgacgg ggtcctcgct tacaaactcc tggtgcagac tggcagccga
     3541 gacaagccca tcgacattag tcagttgact aaacagcgtc tggtagacgc agatggcatc
     3601 attaatccga gcgctttcta catctacctg accgcttggg tcagcaacga ccctgtagct
     3661 tacgctgcct cccaggccaa catccggcct caccggccgg agtgggtcca tgacaaagcc
     3721 gactacatgc cagagaccag gctgagaatc ccagcagcag agcccatcga gtacgctcag
     3781 ttccctttct acctcaacgg cctacgagac acctcagact ttgtggaagc catagaaaaa
     3841 gtgagagtca tctgtaacaa ctatacgagc ctgggactgt ccagctaccc caatggctac
     3901 cccttcctgt tctgggagca atacatcagc ctgcgccact ggctgctgct atccatcagc
     3961 gtggtgctgg cctgcacgtt tctagtgtgc gcagtcttcc tcctgaaccc ctggacggcc
     4021 gggatcattg tcatggtcct ggctctgatg accgttgagc tctttggcat gatgggcctc
     4081 attgggatca agctgagtgc tgtgcctgtg gtcatcctga ttgcatctgt tggcatcgga
     4141 gtggagttca ccgtccacgt ggctttggcc tttctgacag ccattgggga caagaaccac
     4201 agggctatgc tcgctctgga acacatgttt gctcccgttc tggacggtgc tgtgtccact
     4261 ctgctgggtg tactgatgct tgcagggtcc gaatttgatt tcattgtcag atacttcttt
     4321 gccgtcctgg ccattctcac cgtcttgggg gttctcaatg gactggttct gctgcctgtc
     4381 ctcttatcct tctttggacc gtgtcctgag gtgtctccag ccaatggcct aaaccgactg
     4441 cccactcctt cgcctgagcc gcctccaagt gtcgtccggt ttgccgtgcc tcctggtcac
     4501 acgaacaatg ggtctgattc ctccgactcg gagtacagct ctcagaccac ggtgtctggc
     4561 atcagtgagg agctcaggca atacgaagca cagcagggtg ccggaggccc tgcccaccaa
     4621 gtgattgtgg aagccacaga aaaccctgtc tttgcccggt ccactgtggt ccatccggac
     4681 tccagacatc agcctccctt gacccctcgg caacagcccc acctggactc tggctccttg
     4741 tcccctggac ggcaaggcca gcagcctcga agggatcccc ctagagaagg cttgcggcca
     4801 cccccctaca gaccgcgcag agacgctttt gaaatttcta ctgaagggca ttctggccct
     4861 agcaataggg accgctcagg gccccgtggg gcccgttctc acaaccctcg gaacccaacg
     4921 tccaccgcca tgggcagctc tgtgcccagc tactgccagc ccatcaccac tgtgacggct
     4981 tctgcttcgg tgactgttgc tgtgcatccc ccgcctggac ctgggcgcaa cccccgaggg
     5041 gggccctgtc caggctatga gagctaccct gagactgatc acggggtatt tgaggatcct
     5101 catgtgcctt ttcatgtcag gtgtgagagg agggactcaa aggtggaggt catagagcta
     5161 caggacgtgg aatgtgagga gaggccgtgg gggagcagct ccaactagtc tagagggccc
     5221 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     5281 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5341 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5401 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     5461 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     5521 ctctaggggg tatccccacg cgccctacct ctagctagag cttggcgtaa tcatggtcat
     5581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5641 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
