LOCUS       sequence-378906-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Hanaoka K, Qian F, Boletta A, Bhunia AK, Piontek K, Tsiokas L, 
            Sukhatme VP, Guggino WB, Germino GG
  TITLE     Co-assembly of polycystin-1 and -2 produces unique cation-permeable 
            currents.
  JOURNAL   Nature.   . 408(6815):990-4.
  PUBMED    11140688
REFERENCE   2  (bases 1 to 7483)
  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: "Nature.   .
            408(6815):990-4."
COMMENT     Sequence Label: M-PKD2 (OF2-3)
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..132
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             142..171
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             181..210
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             220..249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             259..288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             322..351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     polyA_signal    3535..3669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3559..3578)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3613..3632
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3782..3802)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3782..3800)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3821..3837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3821..3837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3834..3856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3845..3861
                     /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(3869..3875,3876..3893,3894..3899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3914..3935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4052..4069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4223..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4303..4322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4982..5773,5774..5842))
                     /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     5605..5624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5843..5947)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5973..6428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6110..6131)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6322..6341
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6499..7483
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggagcaaaag ctcatttctg aagaggactt gaattcaagg
      361 gtaccgccag tgaccgcgat ggtgaactcc agtcgcgtgc agcctcagca gcccggggac
      421 gccaagcggc cgcccgcgcc ccgcgcgccg gacccaggcc ggctgatggc tggctgcgcg
      481 gccgtgggcg ccagcctcgc cgccccgggc ggcctctgcg agcagcgggg cctggagatc
      541 gagatgcagc gcatccggca ggcggccgcg cgggaccccc cggccggagc cgcggcctcc
      601 ccttctcctc cgctctcgtc gtgctcccgg caggcgtgga gccgcgataa ccccggcttc
      661 gaggccgagg aggaggagga ggaggtggaa ggggaagaag gcggaatggt ggtggagatg
      721 gacgtagagt ggcgcccggg cagccggagg tcggccgcct cctcggccgt gagctccgtg
      781 ggcgcgcgga gccgggggct tgggggctac cacggcgcgg gccacccgag cgggaggcgg
      841 cgccggcgag aggaccaggg cccgccgtgc cccagcccag tcggcggcgg ggacccgctg
      901 catcgccacc tccccctgga agggcagccg ccccgagtgg cctgggcgga gaggctggtt
      961 cgcgggctgc gaggtctctg gggaacaaga ctcatggagg aaagcagcac taaccgagag
     1021 aaatacctta aaagtgtttt acgggaactg gtcacatacc tcctttttct catagtcttg
     1081 tgcatcttga cctacggcat gatgagctcc aatgtgtact actacacccg gatgatgtca
     1141 cagctcttcc tagacacccc cgtgtccaaa acggagaaaa ctaactttaa aactctgtct
     1201 tccatggaag acttctggaa gttcacagaa ggctccttat tggatgggct gtactggaag
     1261 atgcagccca gcaaccagac tgaagctgac aaccgaagtt tcatcttcta tgagaacctg
     1321 ctgttagggg ttccacgaat acggcaactc cgagtcagaa atggatcctg ctctatcccc
     1381 caggacttga gagatgaaat taaagagtgc tatgatgtct actctgtcag tagtgaagat
     1441 agggctccct ttgggccccg aaatggaacc gcttggatct acacaagtga aaaagacttg
     1501 aatggtagta gccactgggg aatcattgca acttatagtg gagctggcta ttatctggat
     1561 ttgtcaagaa caagagagga aacagctgca caagttgcta gcctcaagaa aaatgtctgg
     1621 ctggaccgag gaaccagggc aacttttatt gacttctcag tgtacaacgc caacattaac
     1681 ctgttctgtg tggtcaggtt attggttgaa ttcccagcaa cagttggtgt gattccatct
     1741 tggcaatttc agcctttaaa gctgatccga tatgtcacaa cttttgattt cttcctggca
     1801 gcctgtgaga ttatcttttg tttctttatc ttttactatg tggtggaaga gatattggaa
     1861 attcgcattc acaaactaca ctatttcagg agtttctgga attgtctgga tgttgtgatc
     1921 gttgtgctgt cagtggtagc tataggaatt aacatataca gaacatcaaa tgtggaggtg
     1981 ctactacagt ttctggaaga tcaaaatact ttccccaact ttgagcatct ggcatattgg
     2041 cagatacagt tcaacaatat agctgctgtc acagtatttt ttgtctggat taagctcttc
     2101 aaattcatca attttaacag gaccatgagc cagctctcga caaccatgtc tcgatgtgcc
     2161 aaagacctgt ttggctttgc tattatgttc ttcattattt tcctagcgta tgctcagttg
     2221 gcataccttg tctttggcac tcaggtcgat gacttcagta ctttccaaga gtgtatcttc
     2281 actcaattcc gtatcatttt gggcgatatc aactttgcag agattgagga agctaatcga
     2341 gttttgggac caatttattt cactacattt gtgttcttta tgttcttcat tcttttgaat
     2401 atgtttttgg ctatcatcaa tgatacttac tctgaagtga aatctgactt ggcacagcag
     2461 aaagctgaaa tggaactctc agatcttatc agaaagggct accataaagc tttggtcaaa
     2521 ctaaaactga aaaaaaatac cgtggatgac atttcagaga gtctgcggca aggaggaggc
     2581 aagttaaact ttgacgaact tcgacaagat ctcaaaggga agggccatac tgatgcagag
     2641 attgaggcaa tattcacaaa gtacgaccaa gatggagacc aagaactgac cgaacatgaa
     2701 catcagcaga tgagagacga cttggagaaa gagagggagg acctggattt ggatcacagt
     2761 tctttaccac gtcccatgag cagccgaagt ttccctcgaa gcctggatga ctctgaggag
     2821 gatgacgatg aagatagcgg acatagctcc agaaggaggg gaagcatttc tagtggcgtt
     2881 tcttacgaag agtttcaagt cctggtgaga cgagtggacc ggatggagca ttccatcggc
     2941 agcatagtgt ccaagattga cgccgtgatc gtgaagctag agattatgga gcgagccaaa
     3001 ctgaagagga gggaggtgct gggaaggctg ttggatgggg tggccgagga tgaaaggctg
     3061 ggtcgtgaca gtgaaatcca tagggaacag atggaacggc tagtacgtga agagttggaa
     3121 cgctgggaat ccgatgatgc agcttcccag atcagtcatg gtttaggcac gccagtggga
     3181 ctaaatggtc aacctcgccc cagaagctcc cgcccatctt cctcccaatc tacagaaggc
     3241 atggaaggtg caggtggaaa tgggagttct aatgtccacg tatgatatgt gtgtttcagt
     3301 atgtgtgttt ctaataagtg aggaagtggc tgtcctgaat tgctgtaaca agcacactat
     3361 ttatatgccc tgaccaccat aggatgctag tctttgtgac cgattgctaa tcttctgcac
     3421 tttaatttat tttatataaa ctttacccat ggttcaaann nnaaaaaaaa aaaaaaaaaa
     3481 aaaagggcgg ccgctcgagc atgcatctag aactatagtg agtcgtatta cgtagatcca
     3541 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3601 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3661 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     3721 gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc agcttttgtt
     3781 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3841 gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata aagtgtaaag
     3901 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3961 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5581 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5941 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6001 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6061 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6121 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6181 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6241 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6301 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6361 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6421 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6481 ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa
     6541 ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa
     6601 ttcaatatgg tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa
     6661 ttcaatatgg cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca
     6721 agtttgagga ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca
     6781 ataatggcgg ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc
     6841 cctatgccaa tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag
     6901 tattccatat atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata
     6961 taccatatat ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc
     7021 tctatatatg gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc
     7081 tcccccattg acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa
     7141 taggaccacc caccattgac gtcaatggga tggctcattg cccattcata tccgttctca
     7201 cgccccctat tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt
     7261 aatagtaact tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc
     7321 ccattgacgt caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg
     7381 tcaatgggga ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg
     7441 tgcctaatgg gaggtctata taagcaatgc tcgtttaggg aac
//
