LOCUS       sequence-238037-        5964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged human NPHP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5964)
  AUTHORS   Hoque M, Chen D, Hess RA, Li FQ, Takemaru KI
  TITLE     CEP164 is essential for efferent duct multiciliogenesis and male 
            fertility.
  JOURNAL   Reproduction. 2021 Jul 8;162(2):129-139. doi: 10.1530/REP-21-0042.
  PUBMED    34085951
REFERENCE   2  (bases 1 to 5964)
  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"; journalName: 
            "Reproduction"; date: "2021-07-8- 8"; volume: "162"; issue: "2"; 
            pages: "129-139"
COMMENT     Sequence Label: HA-hNPHP1
FEATURES             Location/Qualifiers
     source          1..5964
                     /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             92..118
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1992..2011)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1993..2011)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2016..2150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2040..2059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2094..2113
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2263..2283)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2263..2281)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2302..2318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2302..2318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2315..2337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2326..2342
                     /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(2350..2356,2357..2374,2375..2380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2395..2416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2533..2550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2704..3292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2784..2803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3463..4254,4255..4323))
                     /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     4086..4105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4324..4428)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4454..4909)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4591..4612)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4803..4822
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4980..5964
                     /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 ataccatgcg atacccatat gatgtgccag attacgctgg
      121 atccatgctg gcgagacgac agcgagatcc tctccaggcc ctgcggcgcc gcaatcagga
      181 gctgaagcaa caggttgata gtttgctttc tgagagccaa ctgaaagaag ctctagaacc
      241 caataaaaga caacatattt atcaaagagt tggggcacct actgaagaag aggaagaaag
      301 tgaaagtgaa gatagtgaag acagtggtgg ggaggaagaa gatgcagagg aggaagagga
      361 agagaaagag gaaaatgaat ctcacaaatg gtcaaccggt gaagaataca tcgctgttgg
      421 agattttact gctcagcaag ttggagatct tacatttaag aaaggggaaa ttctccttgt
      481 aattgaaaaa aaacctgatg gttggtggat agctaaggat gccaaaggaa atgaaggtct
      541 tgttcccaga acctacctag agccttatag tgaagaagaa gaaggccaag agtcaagtga
      601 agagggcagt gaagaagatg tagaggcggt ggatgaaaca gcagatggag cagaagttaa
      661 gcaaagaact gatccccact ggagtgctgt tcagaaagcg atttcagaga taaacactgt
      721 tgatgtgtta actacgatgg gagctattcc tgcagggttc aggccttcca cgctctcaca
      781 gcttctggag gaagggaatc aatttcgagc aaattacttc ttacaaccag agctcatgcc
      841 ttcacaactg gccttcagag atctgatgtg ggatgctaca gaaggcacta ttaggtcgag
      901 accaagtcgt atttcattga ttctgacatt atggagctgt aaaatgattc ctcttccagg
      961 aatgagcata caggttctca gcagacatgt acgcctctgt ctatttgatg gtaataaggt
     1021 tctgagcaac attcatacag tcagagccac atggcaacct aaaaagccca aaacatggac
     1081 cttttctccc caggttactc gcatcttacc atgtttgctt gatggtgatt gctttatcag
     1141 gtctaattct gcatctccag atcttggaat attatttgaa cttggaattt cttatattcg
     1201 caattcaact ggtgaaagag gagagttaag ctgtggctgg gtgtttctta aactttttga
     1261 tgccagtgga gttcctattc cagcaaaaac ttatgagctt ttcttgaatg gtggtactcc
     1321 ttatgaaaaa ggtattgaag tggacccttc aatatccaga agagcacacg gcagtgtttt
     1381 ctaccagatt atgacaatga gaaggcagcc tcaacttcta gtgaaactga gatccttgaa
     1441 cagaagatca agaaatgtac taagtctact gccagaaaca ttaattggaa atatgtgttc
     1501 tattcacttg ttgatatttt atcgacaaat tcttggagat gtgctcctga aagacaggat
     1561 gagcttgcaa agtactgatt taattagcca tcccatgctg gccaccttcc ccatgctctt
     1621 ggagcagcct gatgtgatgg atgctctcag gagttcgtgg gctggaaaag aaagcacatt
     1681 aaaaagatca gagaagagag acaaagagtt cctgaagtcc acgtttctcc tggtttacca
     1741 tgactgcgtg ctcccacttc tccactccac acgcctaccc ccattcaggt gggcagaaga
     1801 agagactgag actgcacggt ggaaagttat cactgacttc cttaagcaaa accaagaaaa
     1861 ccagggcgcc ctccaagctc tgctgtcacc agacggagtt catgaacctt ttgacctttc
     1921 agagcagacc tatgacttct tgggtgaaat gagaaagaat gcagtgtgac tcgagcctct
     1981 agattctgca gccctatagt gagtcgtatt acgtagatcc agacatgata agatacattg
     2041 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2101 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2161 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg
     2221 ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat
     2281 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2341 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
     2401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4441 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     4501 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     4561 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     4621 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     4681 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     4741 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     4801 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     4861 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     4921 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4981 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
     5041 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
     5101 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
     5161 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
     5221 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
     5281 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
     5341 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
     5401 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
     5461 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
     5521 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
     5581 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
     5641 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
     5701 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
     5761 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
     5821 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
     5881 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     5941 ataagcaatg ctcgtttagg gaac
//
