LOCUS       sequence-164703-        6393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PC1-CTp30 expression cassette with a patient mutation at R4150
            and a GST-tag (850).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6393)
  AUTHORS   Doerr N, Wang Y, Kipp KR, Liu G, Benza JJ, Pletnev V, Pavlov TS, 
            Staruschenko A, Mohieldin AM, Takahashi M, Nauli SM, Weimbs T
  TITLE     Regulation of Polycystin-1 Function by Calmodulin Binding.
  JOURNAL   PLoS One. 2016 Aug 25;11(8):e0161525. doi: 
            10.1371/journal.pone.0161525. eCollection 2016.
  PUBMED    27560828
REFERENCE   2  (bases 1 to 6393)
  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: "PLoS One.";
            date: "2016-08-25"; volume: "11(8):e0161525. doi"; pages: " 
            10.1371/journal.pone.0161525. eCollection 2016"
COMMENT     Sequence Label: pcDNA4/TO h-PKD1 FLS R4150C-GST
FEATURES             Location/Qualifiers
     source          1..6393
                     /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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1635..2288
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2246..2268
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2315..2344
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2360..2377
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2400..2417)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2406..2630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2676..3104
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2763..2782)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2973..2994
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3113..3133)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3118..3448
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3299..3434
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3361..3380
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3496..3543
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3562..3936
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4066..4187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4103..4122)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4157..4176
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4236..4252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4260..4276
                     /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(4284..4290,4291..4308,4309..4314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4467..4484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4638..5226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4718..4737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5397..6188,6189..6257))
                     /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     6020..6039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6258..6362)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 ttttggaacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caatagggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgttat ggcccgctgg cgctaccacg ctttgcgtgg agagctgtac cggccggcct
     1081 gggagcccca ggactacgag atggtggagt tgttcctgcg caggctgcgc ctctggatgg
     1141 gcctcagcaa ggtcaaggag ttctgccaca aagtccgctt tgaagggatg gagccgctgc
     1201 cctctcgctc ctccaggggc tccaaggtat ccccggatgt gcccccaccc agcgctggct
     1261 ccgatgcctc gcacccctcc acctcctcca gccagctgga tgggctgagc gtgagcctgg
     1321 gccggctggg gacaaggtgt gagcccgagc cctcccgcct ccaagccgtg ttcgaggccc
     1381 tgctcaccca gtttgaccga ctcaaccagg ccacagagga cgtctaccag ctggagcagc
     1441 agctgcacag cctgcaaggc cgcaggagca gccgggcgcc cgccggatct tcccgtggcc
     1501 catccccggg cctgcggcca gcactgccca gccgccttgc ccgggccagt cggggtgtgg
     1561 acctggccac tggccccagc aggacacccc ttcgggccaa gaacaaggtc caccccagca
     1621 gcactgcggc cgctatgtcc cctatactag gttattggaa aattaagggc cttgtgcaac
     1681 ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg tatgagcgcg
     1741 atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt cccaatcttc
     1801 cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata cgttatatag
     1861 ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt tcaatgcttg
     1921 aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt aaagactttg
     1981 aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg ttcgaagatc
     2041 gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac ttcatgttgt
     2101 atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg ttcccaaaat
     2161 tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac ttgaaatcca
     2221 gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt ggcgaccatc
     2281 ctccaaaatc ggattagctc gaggtcaccc attcgaacaa aaactcatct cagaagagga
     2341 tctgaatatg cataccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     2401 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2461 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2521 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2581 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2641 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2701 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2761 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2821 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2881 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2941 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3001 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta
     3061 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3121 tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat
     3181 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3241 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3301 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3361 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     3421 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     3481 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     3541 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     3601 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     3661 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     3721 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     3781 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     3841 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     3901 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     3961 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4021 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4081 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4141 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4201 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4441 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4501 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4561 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4621 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4681 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4741 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4801 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4861 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4921 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4981 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5041 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5101 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5161 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5221 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5281 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5341 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5401 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5461 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5521 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5581 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5641 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5701 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5761 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5821 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5881 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5941 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6001 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6061 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6121 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6181 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6241 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6301 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6361 cgcacatttc cccgaaaagt gccacctgac gtc
//
