LOCUS       sequence-413219-        9831 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 9831)
  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 9831)
  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: pMx/ATP11C(E184Q)-HA
FEATURES             Location/Qualifiers
     source          1..9831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(130..150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        135..464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      315..450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             821..1615
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(875..894)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1485..1504
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             1808..2277
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2407..2424)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2578..3166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2658..2677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4191..4295)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4325..4918
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4981..5338
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5397..5813
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5743..5765
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5781..5797
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    5823..6197
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     6135..6151
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    6241..6265
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(9376..9387)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     protein_bind    complement(9710..9734)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             9766..9792
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 accattacca gttggtctgg tgtcaaaaat aataataacc gggcaggcca tgtctgcccg
       61 tatttcgcgt aaggaaatcc attatgtact atttaaactc gagcggccgc cagcacagtg
      121 gtcgactgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt cacgctgccg
      481 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
      541 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
      601 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
      661 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
      721 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
      781 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      841 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      901 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      961 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1021 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1081 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1141 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1201 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1261 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1321 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1381 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1441 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1501 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1561 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1621 actctggggt tcgcatcgtc gacgataaaa taaaagattt tatttagtct ccagaaaaag
     1681 gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc
     1741 aaggcatgga aaaatacata actgagaata gagaagttca gatcaaggtc aggaacagat
     1801 ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     1861 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     1921 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
     1981 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     2041 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     2101 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc
     2161 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     2221 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcacat
     2281 gcagcatgta tcaaaattaa tttggttttt tttcttaagt atttacatta aatggccata
     2341 gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2401 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2461 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2521 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2581 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2641 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2701 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2761 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2821 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2881 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2941 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3001 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3061 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3121 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3181 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3241 gagattatca aaaaggatct tcacctagat ccttttgcgg ccggccgcaa atcaatctaa
     3301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4321 gcataatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     4381 aggcatggaa aaatacataa ctgagaatag aaaagttcag atcaaggtca ggaacagatg
     4441 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4501 ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     4561 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt
     4621 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
     4681 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
     4741 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgatt gactgagtcg
     4801 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
     4861 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
     4921 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
     4981 aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat
     5041 gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     5101 tgacgagttc ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg
     5161 tttttgtggc ccgacctgag tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc
     5221 cctaatagga gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc
     5281 cgtctgaatt tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca
     5341 gcatcgttct gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc
     5401 agactgttac cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg
     5461 ctcacaacca gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat
     5521 ggccaacctt taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc
     5581 aggttaagat caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca
     5641 tcgtgacctg ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc
     5701 ctaagcctcc gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt
     5761 cgaccccgcc tcgatcctcc ctttatccag ccctcactcc ttctctaggc gcccccatat
     5821 ggccatatga gatcttatat ggggcacccc cgccccttgt aaacttccct gaccctgaca
     5881 tgacaagagt tactaacagc ccctctctcc aagctcactt acaggctctc tacttagtcc
     5941 agcacgaagt ctggagacct ctggcggcag cctaccaaga acaactggac cgaccggtgg
     6001 tacctcaccc ttaccgagtc ggcgacacag tgtgggtccg ccgacaccag actaagaacc
     6061 tagaacctcg ctggaaagga ccttacacag tcctgctgac cacccccacc gccctcaaag
     6121 tagacggcat cgcagcttgg atacacgccg cccacgtgaa ggctgccgac cccgggggtg
     6181 gaccatcctc tagactgccg gatctagcta gttaattaag gatctctagt taagctatca
     6241 acaagtttgt acaaaaaagc aggctcttta aaggaaccaa ttcagtcgac tggatccacc
     6301 atgcagatgg tcccatctct ccctccagcc tctgagtgtg ctggagaaga gaaacgagtt
     6361 ggcacacgca cagtgtttgt tggcaatcat ccagtttcgg aaacagaagc ttacattgca
     6421 caaagatttt gtgataatag aatagtctca tctaagtata cactttggaa ttttctccca
     6481 aagaatctgt ttgaacagtt tagaagaatt gcaaattttt attttctcat aatcttcctt
     6541 gtacaggtca cagtagacac accaactagc ccagttacca gtggacttcc acttttcttt
     6601 gttataactg ttacagccat caagcaggga tatgaggatt ggctgagaca cagagctgac
     6661 aatgaagtca acaaaagcac tgtttacatt attgaaaatg caaagcgagt gagaaaagaa
     6721 agtgaaaaaa tcaaggttgg tgatgtagta gaagtacagg cagatgaaac ctttccctgt
     6781 gatcttattc ttctatcatc ttgcaccact gatggaacct gttatgtcac tacagccagt
     6841 cttgatgggc aatccaattg caagacacat tatgcagtac gtgataccat tgcactgtgt
     6901 acagcagaat ccatcgatac cctccgagca gcaattgaat gtgaacagcc tcaacctgac
     6961 ctctacaaat ttgttgggcg aatcaatatc tacagtaata gtcttgaggc tgttgccagg
     7021 tctttgggac ctgaaaatct cttgctgaaa ggagctacgc taaaaaatac cgagaagata
     7081 tatggagttg ctgtttacac tggaatggaa accaaaatgg ctttgaacta ccaagggaaa
     7141 tctcagaaac gttctgctgt tgaaaaatct attaatgctt tcctgattgt atatttattt
     7201 atcttactga ccaaagctgc agtatgcact actctaaagt atgtttggca aagtacccca
     7261 tacaatgatg aaccttggta taaccaaaag actcagaaag agcgagagac cttgaaggtt
     7321 ttaaaaatgt tcaccgactt cctatcattt atggttctat tcaactttat cattcctgtc
     7381 tccatgtacg tcacagtaga aatgcagaaa ttcttgggct ccttcttcat ctcatgggat
     7441 aaggactttt atgatgaaga aattaatgaa ggagccctgg ttaacacatc agaccttaat
     7501 gaagaacttg gtcaggtgga ttatgtattt acagataaga ctggaacact cactgaaaac
     7561 agcatggaat tcattgaatg ctgcatagat ggccacaaat ataaaggtgt aactcaagag
     7621 gttgatggat tatctcaaac tgatggaact ttaacatatt ttgacaaagt agataagaat
     7681 cgagaagagc tgtttctacg tgccttgtgt ttatgtcata ctgtagaaat caaaacaaac
     7741 gatgctgttg atggagctac agaatcagct gaattaacct atatctcctc ttcaccagat
     7801 gaaatagctt tggtgaaagg agctaaaagg tacgggttca catttttagg aaatcgaaat
     7861 ggatatatga gagtagagaa ccaaagaaaa gaaatagaag aatatgaact tcttcacacc
     7921 ttaaactttg atgctgtccg gcgacgtatg agtgtaattg tgaagactca agaaggagac
     7981 atacttctct tttgtaaagg agcagactcg gcagtttttc ccagagtgca aaatcatgaa
     8041 attgagttaa ctaaagtcca tgtggaacgt aatgcaatgg atgggtatcg gacactctgt
     8101 gtagccttca aagaaattgc tccagatgat tatgaaagaa ttaacagaca gctcatagag
     8161 gcaaaaatgg ccttacaaga cagagaagaa aaaatggaaa aagttttcga tgatattgag
     8221 acaaacatga atttaattgg agccactgca gttgaagaca agctacaaga tcaagctgca
     8281 gagaccattg aagctctgca tgcagcaggc ctgaaagtct gggtgctcac tggggacaag
     8341 atggagacag ctaaatccac atgctatgcc tgccgccttt tccagaccaa cactgagctc
     8401 ttagaactaa ccacaaaaac cattgaagaa agtgaaagga aagaagatcg attacatgaa
     8461 ttattgatag aatatcgcaa gaaattgctg catgagtttc ctaaaagtac tagaagcttt
     8521 aaaaaagcat ggacagaaca tcaggaatat ggattaatca tagatggctc cacattgtca
     8581 ctcatactaa attctagtca agactctagt tcaaacaatt acaaaagcat tttcctacaa
     8641 atatgtatga agtgtactgc agtgctctgc tgtcggatgg caccattaca gaaagcccag
     8701 attgtcagaa tggtgaagaa tttaaaaggc agcccaataa ctctgtcgat aggtgatggt
     8761 gccaatgatg ttagtatgat cttggaatcc catgtgggaa taggtattaa aggcaaagaa
     8821 ggtcgccaag cagctaggaa tagcgattat tctgttccaa agtttaaaca cttaaagaaa
     8881 ctgctgttgg ctcatggaca tctatattat gtgagaatag cacaccttgt acagtacttc
     8941 ttctataaga acctttgttt cattttgcca cagtttttgt accagttctt ctgtggattc
     9001 tcacaacagc cactgtatga tgctgcttac cttacaatgt acaatatctg cttcacatcc
     9061 ttgcccatcc tggcctatag tctactggaa cagcacatca acattgacac tctgacctca
     9121 gatccccgat tgtatatgaa aatttctggc aatgccatgc tacagttggg ccccttctta
     9181 tattggacat ttctggctgc ctttgaaggg acagtgttct tctttgggac ttactttctt
     9241 tttcagactg catccctaga agaaaatgga aaggtatacg gaaactggac ttttggaacc
     9301 attgttttta cagtcttagt attcactgta accctgaagc ttgccttgga tacccgattc
     9361 tggacgtgga taaatcactt tgtgatttgg ggttctttag ccttctatgt atttttctca
     9421 ttcttctggg gaggaattat ttggcctttt ctcaagcaac agagaatgta ttttgtattt
     9481 gcccaaatgc tgtcttctgt atccacatgg ttggctataa ttcttctaat atttatcagc
     9541 ctgttccctg agattcttct gatagtatta aagaatgtaa gaagaagaag tgccaggaga
     9601 aatctgagct gtagaagggc atctgactca ttatccgcca gaccttcagt cagacctctt
     9661 cttttacgaa cattctcaga cgaatctaat gtactctcga gatatctaga cccagctttc
     9721 ttgtacaaag tggttgatct agagggcccg cggttcgaac tcgagtatcc atacgatgta
     9781 ccagactacg cagagatcca ctagttcaat tccagctgag cgccggtcgc t
//
