LOCUS       sequence-233155-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ptch1-B for function assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  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 7232)
  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-B
FEATURES             Location/Qualifiers
     source          1..7232
                     /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(4723..4741)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4724..4741)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4761..4778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4767..4991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5075..5091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5075..5091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5088..5110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5099..5115
                     /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(5123..5129,5130..5147,5148..5153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5168..5189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5306..5323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5477..6065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5557..5576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6236..7027,7028..7096))
                     /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     6859..6878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7097..7201)
                     /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 atgcagtcca ccgttcagct ccgcacagag tatgaccctc acacgcacgt gtactacacc
     2821 accgccgagc cacgctctga gatctctgta cagcctgtta ccgtcaccca ggacaacctc
     2881 agctgtcaga gtcccgagag caccagctct accagggacc tgctctccca gttctcagac
     2941 tccagcctcc actgcctcga gcccccctgc accaagtgga cactctcttc gtttgcagag
     3001 aagcactatg ctcctttcct cctgaaaccc aaagccaagg ttgtggtaat ccttcttttc
     3061 ctgggcttgc tgggggtcag cctttatggg accacccgag tgagagacgg gctggacctc
     3121 acggacattg ttccccggga aaccagagaa tatgacttca tagctgccca gttcaagtac
     3181 ttctctttct acaacatgta tatagtcacc cagaaagcag actacccgaa tatccagcac
     3241 ctactttacg accttcataa gagtttcagc aatgtgaagt atgtcatgct ggaggagaac
     3301 aagcaacttc cccaaatgtg gctgcactac tttagagact ggcttcaagg acttcaggat
     3361 gcatttgaca gtgactggga aactgggagg atcatgccaa acaattataa aaatggatca
     3421 gatgacgggg tcctcgctta caaactcctg gtgcagactg gcagccgaga caagcccatc
     3481 gacattagtc agttgactaa acagcgtctg gtagacgcag atggcatcat taatccgagc
     3541 gctttctaca tctacctgac cgcttgggtc agcaacgacc ctgtagctta cgctgcctcc
     3601 caggccaaca tccggcctca ccggccggag tgggtccatg acaaagccga ctacatgcca
     3661 gagaccaggc tgagaatccc agcagcagag cccatcgagt acgctcagtt ccctttctac
     3721 ctcaacggcc tacgagacac ctcagacttt gtggaagcca tagaaaaagt gagagtcatc
     3781 tgtaacaact atacgagcct gggactgtcc agctacccca atggctaccc cttcctgttc
     3841 tgggagcaat acatcagcct gcgccactgg ctgctgctat ccatcagcgt ggtgctggcc
     3901 tgcacgtttc tagtgtgcgc agtcttcctc ctgaacccct ggacggccgg gatcattgtc
     3961 atggtcctgg ctctgatgac cgttgagctc tttggcatga tgggcctcat tgggatcaag
     4021 ctgagtgctg tgcctgtggt catcctgatt gcatctgttg gcatcggagt ggagttcacc
     4081 gtccacgtgg ctttggcctt tctgacagcc attggggaca agaaccacag ggctatgctc
     4141 gctctggaac acatgtttgc tcccgttctg gacggtgctg tgtccactct gctgggtgta
     4201 ctgatgcttg cagggtccga atttgatttc attgtcagat acttctttgc cgtcctggcc
     4261 attctcaccg tcttgggggt tctcaatgga ctggttctgc tgcctgtcct cttatccttc
     4321 tttggaccgt gtcctgaggt gtctccagcc aatggcctaa accgactgcc cactccttcg
     4381 cctgagccgc ctccaagtgt cgtccggttt gccgtgcctc ctggtcacac gaacaatggg
     4441 tctgattcct ccgactcgga gtacagctct cagaccacgg tgtctggcat cagtgaggag
     4501 ctcaggcaat acgaagcaca gcagggtgcc ggaggccctg cccaccaagt gattgtggaa
     4561 gccacagaaa accctgtctt tgcccggtcc actgtggtcc atccggactc cagacatcag
     4621 cctcccttga cccctcggca acagccccac ctggactctg gctccttgtc ccctggacgg
     4681 caaggccagc agcctcgaag ggattagtct agagggccct attctatagt gtcacctaaa
     4741 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     4801 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     4861 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4921 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     4981 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     5041 gccctacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5101 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5161 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5221 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5281 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5881 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     6961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7201 gcacatttcc ccgaaaagtg ccacctgacg tc
//
