LOCUS       sequence-413149-        9885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATP8A1 mutant E191Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9885)
  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 9885)
  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/ATP8A1(E191Q)-HA
FEATURES             Location/Qualifiers
     source          1..9885
                     /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(5461..5485)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5517..5543
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5712..5732)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5717..6046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5897..6032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6403..7197
                     /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(6457..6476)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7067..7086
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             7390..7859
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7989..8006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8160..8748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8240..8259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8912..9703,9704..9772))
                     /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     9535..9554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9773..9877)
                     /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 ctcactcggc 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 cccaccatgc ggaggaccgt gtcggagatc cgctcgcgcg
     2041 ccgaaggtta tgagaagaca gatgatgttt cagagaagac ctcactggct gaccaggagg
     2101 aagtaaggac tattttcatc aaccagcccc agctgacaaa attctgcaat aaccatgtca
     2161 gcactgcaaa atacaacata atcacattcc ttccaagatt tctctactct cagttcagaa
     2221 gagctgctaa ttcatttttt ctctttattg cactgctgca gcaaatacct gatgtgtcac
     2281 caacaggtcg ttatacaaca ctggttcctc tcttatttat tttagctgtg gcagctatca
     2341 aagagataat agaagatatt aaacgacata aagctgataa tgcagtgaac aagaaacaaa
     2401 cgcaagtttt gagaaatggt gcttgggaaa ttgtccactg ggaaaaggta aatgttggag
     2461 atatagttat aataaaaggc aaagagtata tacctgctga cactgtactt ctctcatcaa
     2521 gtgagcccca agccatgtgc tacattgaaa catccaactt agatggtcaa acaaacttga
     2581 aaattagaca gggcttacca gcaacatcag atatcaaaga cgttgacagt ttgatgagga
     2641 tttctggcag aattgagtgt gaaagtccaa acagacatct ctacgatttt gttggaaaca
     2701 taaggcttga tggacatggc accgttccac tgggagcaga tcagattctt cttcgaggag
     2761 ctcagttgag aaatacacag tgggttcatg gaatagttgt ctacactgga catgacacca
     2821 agctgatgca gaattcaaca agtccaccac ttaagctctc aaatgtggaa cggattacaa
     2881 atgtacaaat tttgatttta ttttgtatct taattgccat gtctcttgtc tgttctgtgg
     2941 gctcagccat ttggaatcga aggcattctg gaaaagactg gtatctcaat ctaaactatg
     3001 gtggcgctag taattttgga ctgaatttct tgaccttcat catccttttc aacaatctca
     3061 ttcctatcag cttattggtt acattagaag ttgtgaaatt tacccaggca tacttcataa
     3121 attgggatct tgacatgcac tatgaaccca cagacactgc tgctatggct cgaacatcta
     3181 atctgaatga ggaacttggc caggttaaat acatattttc tgacaaaact ggtactctga
     3241 catgcaatgt aatgcagttt aagaagtgca ccatagcggg agttgcttat gggcagaact
     3301 cacagtttgg agatgaaaaa acatttagtg attcatcatt gctggaaaat ctccaaaata
     3361 atcatccaac tgcacctata atatgtgaat ttcttacaat gatggcagtc tgtcacacag
     3421 cagtgccaga gcgagaaggt gacaagatta tttatcaagc agcatctcca gatgagggag
     3481 cattggtcag agcagccaag caattgaatt ttgttttcac tggaagaaca cccgactcgg
     3541 tgattataga ttcactgggg caggaagaaa gatatgaatt gctcaatgtc ttggagttta
     3601 ccagtgctag gaaaagaatg tcagtgattg ttcgcactcc atctggaaag ttacgactct
     3661 actgcaaagg agctgacact gtaatttatg atcgactggc agagacgtca aaatacaaag
     3721 aaattaccct aaaacattta gagcagtttg ctacagaagg gttaagaact ttatgttttg
     3781 ctgtggctga gatttcagag agcgactttc aggagtggcg agcagtctat cagcgagcat
     3841 ctacatctgt gcagaacagg ctactcaaac tcgaagagag ttatgagttg attgaaaaga
     3901 atcttcagct acttggagca acagccattg aggataaatt acaagatcaa gtgcctgaaa
     3961 ccatagaaac gctaatgaaa gcagacatca aaatctggat ccttacaggg gacaagcaag
     4021 aaactgccat taacatcgga cactcctgca aactgttgaa gaagaacatg ggaatgattg
     4081 ttataaatga aggctctctt gatggaacaa gggaaactct cagtcgtcac tgtactaccc
     4141 ttggtgatgc tctccggaaa gagaatgatt ttgctcttat aattgatggg aaaaccctca
     4201 aatatgcctt aacctttgga gtacgacagt atttcctgga cttagctttg tcatgcaaag
     4261 ctgtcatttg ctgtcgggtt tctcctcttc aaaaatctga agttgttgag atggttaaga
     4321 aacaagtcaa agtcgtaacg cttgcaatcg gtgatggagc aaatgatgtc agcatgatac
     4381 agacagcgca cgttggtgtt ggtatcagtg gcaatgaagg cctgcaggca gctaattcct
     4441 ctgactactc catagctcag ttcaaatatt tgaagaattt actgatgatt catggtgcct
     4501 ggaactataa cagagtctcc aagtgcatct tatactgctt ctacaagaat atagtgctct
     4561 atattatcga gatctggttt gcctttgtta atggcttttc tggacagatc ctctttgaaa
     4621 gatggtgtat aggtctctat aacgtgatgt ttacagcaat gcctccttta actcttggaa
     4681 tatttgagag atcatgcaga aaagagaaca tgttgaagta ccctgaatta tacaaaacat
     4741 ctcagaatgc cctggacttc aacaccaagg ttttctgggt tcattgttta aatggcctct
     4801 tccactcagt tattctgttt tggtttccac taaaagccct tcagtatggt actgcatttg
     4861 gaaatgggaa aacctcggat tatctgctac tgggaaactt tgtgtacact tttgtggtga
     4921 taactgtgtg tttgaaagct ggattggaga catcatattg gacatggttc agccacatag
     4981 cgatatgggg gagcatcgca ctctgggtgg tgttttttgg aatctactca tctctgtggc
     5041 ctgccattcc gatggcccct gatatgtcag gagaggcagc catgttgttc agttctggag
     5101 tcttttggat gggcttgtta ttcatccctg tggcatctct gctccttgat gtggtgtaca
     5161 aggttatcaa gaggactgct tttaaaacat tggtcgatga agttcaggag ctggaggcaa
     5221 aatctcaaga cccaggagca gttgtacttg gaaaaagcct gaccgagagg gcgcaactgc
     5281 tcaagaacgt ctttaagaag aaccacgtga acttgtaccg ctctgaatcc ttgcaacaaa
     5341 atctgctcca tgggtatgcg ttctctcaag atgaaaatgg aatcgtttca cagtctgaag
     5401 tgataagagc atatgatacc acgaaacaga ggcccgacga atgggcctcg agatatctag
     5461 acccagcttt cttgtacaaa gtggttgatc tagagggccc gcggttcgaa ctcgagtatc
     5521 catacgatgt accagactac gcagagatcc actagttcaa ttccagctga gcgccggtcg
     5581 ctaccattac cagttggtct ggtgtcaaaa ataataataa ccgggcaggc catgtctgcc
     5641 cgtatttcgc gtaaggaaat ccattatgta ctatttaaac tcgagcggcc gccagcacag
     5701 tggtcgactg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5881 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6001 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttcacgctgc
     6061 cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa agccagtccg
     6121 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac
     6181 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg
     6241 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt
     6301 gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg
     6361 ggatcaagat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     6421 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     6481 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     6541 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     6601 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     6661 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6721 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6781 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6841 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6901 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6961 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     7021 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     7081 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     7141 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     7201 ggactctggg gttcgcatcg tcgacgataa aataaaagat tttatttagt ctccagaaaa
     7261 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     7321 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     7381 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     7441 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     7501 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     7561 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     7621 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     7681 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg attgactgag
     7741 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     7801 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     7861 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     7921 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     8041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     8101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggccttt
     8161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     8221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     8281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     8341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     8401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     8461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     8521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     8581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     8641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     8701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8821 atgagattat caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct
     8881 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8941 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9001 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9061 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9841 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttc
//
