LOCUS       sequence-159101-        5728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human PC1-CTp30 expression cassette with a point mutant that 
            disrupts the second calmodulin binding domain (514).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5728)
  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 5728)
  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 S4144D-Myc/His
FEATURES             Location/Qualifiers
     source          1..5728
                     /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"
     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             1650..1679
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1695..1712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1735..1752)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1741..1965
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2011..2439
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2308..2329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2448..2468)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2453..2783
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2634..2769
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2696..2715
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2831..2878
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2897..3271
                     /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    3401..3522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3438..3457)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3492..3511
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3571..3587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3571..3587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3584..3606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3595..3611
                     /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(3619..3625,3626..3643,3644..3649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3664..3685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3802..3819)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3973..4561)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4053..4072)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4732..5523,5524..5592))
                     /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     5355..5374
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5593..5697)
                     /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 caaatgggcg
      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 gcctcgacaa ggtcaaggag ttccgccaca 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 cgctcgaggt cacccattcg aacaaaaact catctcagaa gaggatctga
     1681 atatgcatac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     1741 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     1801 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     1861 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     1921 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     1981 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2161 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2341 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt
     2401 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     2461 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     2521 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2581 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2641 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2701 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     2761 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     2821 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     2881 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     2941 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     3001 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     3061 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     3121 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     3181 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     3241 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     3301 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3361 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     3421 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3481 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     3541 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3601 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3661 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3721 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3781 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3841 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3901 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3961 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4021 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4081 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4141 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     4201 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4261 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4321 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4381 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4441 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4501 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4561 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4621 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4681 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4741 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4801 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4861 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4921 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4981 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5041 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5101 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5161 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5221 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5281 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5341 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5401 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5461 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5521 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5581 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5701 atttccccga aaagtgccac ctgacgtc
//
