LOCUS       sequence-413168-        9840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATP11A mutant E186Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9840)
  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 9840)
  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/ATP11A(E186Q)-HA
FEATURES             Location/Qualifiers
     source          1..9840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1440..1462
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1478..1494
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1832..1848
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     protein_bind    1938..1962
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5416..5440)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5472..5498
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5667..5687)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5672..6001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5852..5987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6358..7152
                     /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(6412..6431)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7022..7041
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             7345..7814
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7944..7961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8115..8703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8195..8214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8867..9658,9659..9727))
                     /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     9490..9509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9728..9832)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ctctagttaa gctatcaaca agtttgtaca aaaaagcagg ctctttaaag gaaccaattc
     1981 agtcgactgg atccaccatg gactgcagcc tcgtgcggac gctcgtgcac agatactgtg
     2041 caggagaaga gaattgggtg gacagcagga ccatctacgt gggacacagg gagccacctc
     2101 cgggcgcaga ggcctacatc ccacagagat acccagacaa caggatcgtc tcgtccaagt
     2161 acacattttg gaactttata cccaagaatt tatttgaaca attcagaaga gtagccaact
     2221 tttatttcct tatcatattt ctggtgcagt tgattattga tacacccaca agtccagtga
     2281 caagcggact tccactcttc tttgtcatta ctgtgacggc tatcaaacag ggttatgaag
     2341 actggcttcg acataaagca gacaatgcca tgaaccagtg tcctgttcat ttcattcagc
     2401 acggcaagct cgttcggaaa caaagtcgaa agctgcgagt tggggacatt gtcatggtta
     2461 aggaggacga gacctttccc tgcgacttga tcttcctttc cagcaaccgg ggagatggga
     2521 cgtgccacgt caccaccgcc agcttggatg gacaatccag ccataaaacg cattacgcag
     2581 tccaggacac caaaggcttc cacacagagg aggatatcgg cggacttcac gccaccatcg
     2641 agtgtgagca gccccagccc gacctctaca agttcgtggg tcgcatcaac gtttacagtg
     2701 acctgaatga ccccgtggtg aggcccttag gatcggaaaa cctgctgctt agaggagcta
     2761 cactgaagaa cactgagaaa atctttggtg tggctattta cacgggaatg gaaaccaaga
     2821 tggcattaaa ttatcaatca aaatctcaga agcgatctgc cgtggaaaaa tcgatgaatg
     2881 cgttcctcat tgtgtatctc tgcattctga tcagcaaagc cctgataaac actgtgctga
     2941 aatacgtgtg gcagagtgag ccctttcggg atgagccgtg gtataatcag aaaacggagt
     3001 cggaaaggca gaggaatctg ttcctcaagg cattcacgga cttcctggcc ttcatggtcc
     3061 tctttaacta catcatccct gtgtccatgt acgtcacggt cgagatgcag aagttcctcg
     3121 gctcttactt catcacctgg gacgaagaca tgtttgacga ggagactggc gaggggcctc
     3181 tggtgaacac gtcggacctc aatgaagagc tgggacaggt ggagtacatc ttcacagaca
     3241 agaccggcac cctcacggaa aacaacatgg agttcaagga gtgctgcatc gaaggccatg
     3301 tctacgtgcc ccacgtcatc tgcaacgggc aggtcctccc agagtcgtca ggaatcgaca
     3361 tgattgactc gtcccccagc gtcaacggga gggagcgcga ggagctgttt ttccgggccc
     3421 tctgtctctg ccacaccgtc caggtgaaag acgatgacag cgtagacggc cccaggaaat
     3481 cgccggacgg ggggaaatcc tgtgtgtaca tctcatcctc gcccgacgag gtggcgctgg
     3541 tcgaaggtgt ccagagactt ggctttacct acctaaggct gaaggacaat tacatggaga
     3601 tattaaacag ggagaaccac atcgaaaggt ttgaattgct ggaaattttg agttttgact
     3661 cagtcagaag gagaatgagt gtaattgtaa aatctgctac aggagaaatt tatctgtttt
     3721 gcaaaggagc agattcttcg atattccccc gagtgataga aggcaaagtt gaccagatcc
     3781 gagccagagt ggagcgtaac gcagtggagg ggctccgaac tttgtgtgtt gcttataaaa
     3841 ggctgatcca agaagaatat gaaggcattt gtaagctgct gcaggctgcc aaagtggccc
     3901 ttcaagatcg agagaaaaag ttagcagaag cctatgagca aatagagaaa gatcttactc
     3961 tgcttggtgc tacagctgtt gaggaccggc tgcaggagaa agctgcagac accatcgagg
     4021 ccctgcagaa ggccgggatc aaagtctggg ttctcacggg agacaagatg gagacggccg
     4081 cggccacgtg ctacgcctgc aagctcttcc gcaggaacac gcagctgctg gagctgacca
     4141 ccaagaggat cgaggagcag agcctgcacg acgtcctgtt cgagctgagc aagacggtcc
     4201 tgcgccacag cgggagcctg accagagaca acctctccgg actttcagca gatatgcagg
     4261 actacggttt aattatcgac ggagctgcac tgtctctgat aatgaagcct cgagaagacg
     4321 ggagttccgg caactacagg gagctcttcc tggaaatctg ccggagctgc agcgcggtgc
     4381 tctgctgccg catggcgccc ttgcagaagg ctcagattgt taaattaatc aaattttcaa
     4441 aagagcaccc aatcacgtta gcaattggcg atggtgcaaa tgatgtcagc atgattctgg
     4501 aagcgcacgt gggcataggt gtcatcggca aggaaggccg ccaggctgcc aggaacagcg
     4561 actatgcaat cccaaagttt aagcatttga agaagatgct gcttgttcac gggcattttt
     4621 attacattag gatctctgag ctcgtgcagt acttcttcta taagaacgtc tgcttcatct
     4681 tccctcagtt tttataccag ttcttctgtg ggttttcaca acagactttg tacgacaccg
     4741 cgtatctgac cctctacaac atcagcttca cctccctccc catcctcctg tacagcctca
     4801 tggagcagca tgttggcatt gacgtgctca agagagaccc gaccctgtac agggacgtcg
     4861 ccaagaatgc cctgctgcgc tggcgcgtgt tcatctactg gacgctcctg ggactgtttg
     4921 acgcactggt gttcttcttt ggtgcttatt tcgtgtttga aaatacaact gtgacaagca
     4981 acgggcagat atttggaaac tggacgtttg gaacgctggt attcaccgtg atggtgttca
     5041 cagttacact aaagcttgca ttggacacac actactggac ttggatcaac cattttgtca
     5101 tctgggggtc gctgctgttc tacgttgtct tttcacttct ctggggagga gtgatctggc
     5161 cgttcctcaa ctaccagagg atgtactacg tgttcatcca gatgctgtcc agcgggcccg
     5221 cctggctggc catcgtgctg ctggtgacca tcagcctcct tcccgacgtc ctcaagaaag
     5281 tcctgtgccg gcagctgtgg ccaacagcaa cagagagagt ccagactaag agccagtgcc
     5341 tttctgtcga gcagtcaacc atctttatgc tttctcagac ttccagcagc ctgagtttcg
     5401 cgtcgagata tctagaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt
     5461 tcgaactcga gtatccatac gatgtaccag actacgcaga gatccactag ttcaattcca
     5521 gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat aataaccggg
     5581 caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt taaactcgag
     5641 cggccgccag cacagtggtc gactgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5701 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5761 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5821 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5881 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5941 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6001 aaagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
     6061 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     6121 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     6181 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     6241 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     6301 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
     6361 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6421 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6481 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6541 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6601 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6661 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6721 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6781 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6841 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6901 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6961 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7021 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7081 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7141 gagttcttct gagcgggact ctggggttcg catcgtcgac gataaaataa aagattttat
     7201 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt
     7261 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     7321 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     7381 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga
     7441 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     7501 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     7561 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     7621 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc
     7681 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     7741 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     7801 gcgggggtct ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt
     7861 tacattaaat ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg
     7921 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     8041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     8101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     8161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     8221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     8281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     8341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     8401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8521 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     8581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg
     8821 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8881 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8941 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9001 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9061 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9121 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9181 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9241 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9301 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9361 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9421 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9481 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9541 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9601 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9661 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9721 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9781 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
//
