LOCUS       sequence-413047-        8863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATP11C mutant E184Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8863)
  AUTHORS   Takatsu H, Tanaka G, Segawa K, Suzuki J, Nagata S, Nakayama K, Shin 
            HW
  TITLE     Phospholipid flippase activities and substrate specificities of 
            human type IV P-type ATPases localized to the plasma membrane.
  JOURNAL   J Biol Chem. 2014 Nov 28;289(48):33543-56. doi: 
            10.1074/jbc.M114.593012. Epub 2014 Oct 14.
  PUBMED    25315773
REFERENCE   2  (bases 1 to 8863)
  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: 
            "10.1074/jbc.M114.593012"; journalName: "J Biol Chem"; date: 
            "2014-11-28- 28"; volume: "289"; issue: "48"; pages: "33543-56"
COMMENT     Sequence Label: pCAG/ATP11C(E184Q)-HA
FEATURES             Location/Qualifiers
     source          1..8863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             400..1203
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(463..482)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1073..1092
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1210..1258
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        1268..1647
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2176..2198
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2260..2279
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2348..2372
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5483..5494)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     protein_bind    complement(5817..5841)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5885..5911
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5999..6018)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6064..6119
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6118..6137)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6480..6496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6480..6496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6493..6515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6504..6520
                     /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(6528..6534,6535..6552,6553..6558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6573..6594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6653..6848
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6699..6834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6854..6988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6891..6910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6945..6964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7056..7073)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7227..7815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7307..7326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7986..8777,8778..8846))
                     /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     8609..8628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8847..8863,1..88))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
       61 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcga
      121 gcagtgtggt tttcaagagg aagcaaaaag cctctccacc caggcctgga atgtttccac
      181 ccaatgtcga gcagtgtggt tttgcaagag gaagcaaaaa gcctctccac ccaggcctgg
      241 aatgtttcca cccaatgtcg agcaaacccc gcccagcgtc ttgtcattgg cgaattaatt
      301 cgaacacgca gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga
      361 cgcgtgtggc ctcgaacacc gagcgaccct gcagccaata tgggatcggc cattgaacaa
      421 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
      481 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
      541 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
      601 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
      661 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
      721 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
      781 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
      841 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
      901 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgatgatctc
      961 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1021 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     1081 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1141 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1201 tgaggggatc ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcgg
     1261 atccgtcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     1321 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     1381 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1441 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1501 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     1561 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     1621 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
     1681 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
     1741 gcagcgatgg gggcgggggg gggggggggg gcggcccggg gggggggggg gggggggggg
     1801 gggggggggg gggggggggg gggggggggg gggggggccc gggggggggg ggggggggcg
     1861 gggggggggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     1921 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     1981 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     2041 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     2101 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     2161 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     2221 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     2281 tgtctcatca ttttggcaaa gaatttgcta gcgagctcgc ggccgcagat ctctagttaa
     2341 gctatcaaca agtttgtaca aaaaagcagg ctctttaaag gaaccaattc agtcgactgg
     2401 atccaccatg cagatggtcc catctctccc tccagcctct gagtgtgctg gagaagagaa
     2461 acgagttggc acacgcacag tgtttgttgg caatcatcca gtttcggaaa cagaagctta
     2521 cattgcacaa agattttgtg ataatagaat agtctcatct aagtatacac tttggaattt
     2581 tctcccaaag aatctgtttg aacagtttag aagaattgca aatttttatt ttctcataat
     2641 cttccttgta caggtcacag tagacacacc aactagccca gttaccagtg gacttccact
     2701 tttctttgtt ataactgtta cagccatcaa gcagggatat gaggattggc tgagacacag
     2761 agctgacaat gaagtcaaca aaagcactgt ttacattatt gaaaatgcaa agcgagtgag
     2821 aaaagaaagt gaaaaaatca aggttggtga tgtagtagaa gtacaggcag atgaaacctt
     2881 tccctgtgat cttattcttc tatcatcttg caccactgat ggaacctgtt atgtcactac
     2941 agccagtctt gatgggcaat ccaattgcaa gacacattat gcagtacgtg ataccattgc
     3001 actgtgtaca gcagaatcca tcgataccct ccgagcagca attgaatgtg aacagcctca
     3061 acctgacctc tacaaatttg ttgggcgaat caatatctac agtaatagtc ttgaggctgt
     3121 tgccaggtct ttgggacctg aaaatctctt gctgaaagga gctacgctaa aaaataccga
     3181 gaagatatat ggagttgctg tttacactgg aatggaaacc aaaatggctt tgaactacca
     3241 agggaaatct cagaaacgtt ctgctgttga aaaatctatt aatgctttcc tgattgtata
     3301 tttatttatc ttactgacca aagctgcagt atgcactact ctaaagtatg tttggcaaag
     3361 taccccatac aatgatgaac cttggtataa ccaaaagact cagaaagagc gagagacctt
     3421 gaaggtttta aaaatgttca ccgacttcct atcatttatg gttctattca actttatcat
     3481 tcctgtctcc atgtacgtca cagtagaaat gcagaaattc ttgggctcct tcttcatctc
     3541 atgggataag gacttttatg atgaagaaat taatgaagga gccctggtta acacatcaga
     3601 ccttaatgaa gaacttggtc aggtggatta tgtatttaca gataagactg gaacactcac
     3661 tgaaaacagc atggaattca ttgaatgctg catagatggc cacaaatata aaggtgtaac
     3721 tcaagaggtt gatggattat ctcaaactga tggaacttta acatattttg acaaagtaga
     3781 taagaatcga gaagagctgt ttctacgtgc cttgtgttta tgtcatactg tagaaatcaa
     3841 aacaaacgat gctgttgatg gagctacaga atcagctgaa ttaacctata tctcctcttc
     3901 accagatgaa atagctttgg tgaaaggagc taaaaggtac gggttcacat ttttaggaaa
     3961 tcgaaatgga tatatgagag tagagaacca aagaaaagaa atagaagaat atgaacttct
     4021 tcacacctta aactttgatg ctgtccggcg acgtatgagt gtaattgtga agactcaaga
     4081 aggagacata cttctctttt gtaaaggagc agactcggca gtttttccca gagtgcaaaa
     4141 tcatgaaatt gagttaacta aagtccatgt ggaacgtaat gcaatggatg ggtatcggac
     4201 actctgtgta gccttcaaag aaattgctcc agatgattat gaaagaatta acagacagct
     4261 catagaggca aaaatggcct tacaagacag agaagaaaaa atggaaaaag ttttcgatga
     4321 tattgagaca aacatgaatt taattggagc cactgcagtt gaagacaagc tacaagatca
     4381 agctgcagag accattgaag ctctgcatgc agcaggcctg aaagtctggg tgctcactgg
     4441 ggacaagatg gagacagcta aatccacatg ctatgcctgc cgccttttcc agaccaacac
     4501 tgagctctta gaactaacca caaaaaccat tgaagaaagt gaaaggaaag aagatcgatt
     4561 acatgaatta ttgatagaat atcgcaagaa attgctgcat gagtttccta aaagtactag
     4621 aagctttaaa aaagcatgga cagaacatca ggaatatgga ttaatcatag atggctccac
     4681 attgtcactc atactaaatt ctagtcaaga ctctagttca aacaattaca aaagcatttt
     4741 cctacaaata tgtatgaagt gtactgcagt gctctgctgt cggatggcac cattacagaa
     4801 agcccagatt gtcagaatgg tgaagaattt aaaaggcagc ccaataactc tgtcgatagg
     4861 tgatggtgcc aatgatgtta gtatgatctt ggaatcccat gtgggaatag gtattaaagg
     4921 caaagaaggt cgccaagcag ctaggaatag cgattattct gttccaaagt ttaaacactt
     4981 aaagaaactg ctgttggctc atggacatct atattatgtg agaatagcac accttgtaca
     5041 gtacttcttc tataagaacc tttgtttcat tttgccacag tttttgtacc agttcttctg
     5101 tggattctca caacagccac tgtatgatgc tgcttacctt acaatgtaca atatctgctt
     5161 cacatccttg cccatcctgg cctatagtct actggaacag cacatcaaca ttgacactct
     5221 gacctcagat ccccgattgt atatgaaaat ttctggcaat gccatgctac agttgggccc
     5281 cttcttatat tggacatttc tggctgcctt tgaagggaca gtgttcttct ttgggactta
     5341 ctttcttttt cagactgcat ccctagaaga aaatggaaag gtatacggaa actggacttt
     5401 tggaaccatt gtttttacag tcttagtatt cactgtaacc ctgaagcttg ccttggatac
     5461 ccgattctgg acgtggataa atcactttgt gatttggggt tctttagcct tctatgtatt
     5521 tttctcattc ttctggggag gaattatttg gccttttctc aagcaacaga gaatgtattt
     5581 tgtatttgcc caaatgctgt cttctgtatc cacatggttg gctataattc ttctaatatt
     5641 tatcagcctg ttccctgaga ttcttctgat agtattaaag aatgtaagaa gaagaagtgc
     5701 caggagaaat ctgagctgta gaagggcatc tgactcatta tccgccagac cttcagtcag
     5761 acctcttctt ttacgaacat tctcagacga atctaatgta ctctcgagat atctagaccc
     5821 agctttcttg tacaaagtgg ttgatctaga gggcccgcgg ttcgaactcg aggtaccgaa
     5881 ttcgtaccca tacgacgtcc cagactacgc ttaattcact cctcaggtgc aggctgccta
     5941 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatcgatc
     6001 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     6061 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     6121 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     6181 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     6241 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     6301 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     6361 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     6421 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     6481 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6541 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6601 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     6661 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6721 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6781 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6841 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6901 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6961 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     7021 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7081 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7141 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7201 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7261 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7321 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7381 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7441 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7501 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7561 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7621 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7681 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7741 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7801 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7861 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7921 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7981 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8041 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8101 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8161 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8221 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8281 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8341 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8401 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8461 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8521 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8581 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8641 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8701 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8761 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8821 gcgacacgga aatgttgaat actcatactc ttcctttttc aat
//
