LOCUS       sequence-413482-        9966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ATP11B mutant E180Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9966)
  AUTHORS   Tone T, Nakayama K, Takatsu H, Shin HW
  TITLE     ATPase reaction cycle of P4-ATPases affects their transport from the
            endoplasmic reticulum.
  JOURNAL   FEBS Lett. 2020 Feb;594(3):412-423. doi: 10.1002/1873-3468.13629. 
            Epub 2019 Oct 14.
  PUBMED    31571211
REFERENCE   2  (bases 1 to 9966)
  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: 
            "10.1002/1873-3468.13629"; journalName: "FEBS Lett"; date: 
            "2020-02"; volume: "594"; issue: "3"; pages: "412-423"
COMMENT     Sequence Label: pMx/ATP11B(E180Q)-HA
FEATURES             Location/Qualifiers
     source          1..9966
                     /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(5542..5566)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5598..5624
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5793..5813)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5798..6127
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5978..6113
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6484..7278
                     /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(6538..6557)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7148..7167
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             7471..7940
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(8070..8087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8241..8829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8321..8340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8993..9784,9785..9853))
                     /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     9616..9635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9854..9958)
                     /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 atctaccatg tggcgctgga tccggcagca gctgggtttt gacccaccac
     2041 atcagagtga cacaagaacc atctacgtag ccaacaggtt tcctcagaat ggcctttaca
     2101 cacctcagaa atttatagat aacaggatca tttcatctaa gtacactgtg tggaattttg
     2161 ttccaaaaaa tttatttgaa cagttcagaa gagtggcaaa cttttatttt cttattatat
     2221 ttttggttca gcttatgatt gatacaccta ccagtccagt taccagtgga cttccattat
     2281 tctttgtgat aacagtaact gccataaagc agggatatga agattggtta cggcataact
     2341 cagataatga agtaaatgga gctcctgttt atgttgttcg aagtggtggc cttgtaaaaa
     2401 ctagatcaaa aaacattcgg gtgggtgata ttgttcgaat agccaaagat gaaatttttc
     2461 ctgcagactt ggtgcttctg tcctcagatc gactggatgg ttcctgtcac gttacaactg
     2521 ctagtttgga cggacaaact aacctgaaga cacatgtggc agttccagaa acagcattat
     2581 tacaaacagt tgccaatttg gacactctag tagctgtaat agaatgccag caaccagaag
     2641 cagacttata cagattcatg ggacgaatga tcataaccca acaaatggaa gaaattgtaa
     2701 gacctctggg gccggagagt ctcctgcttc gtggagccag attaaaaaac acaaaagaaa
     2761 tttttggtgt tgcggtatac actggaatgg aaactaagat ggcattaaat tacaagagca
     2821 aatcacagaa acgatctgca gtagaaaagt caatgaatac atttttgata atttatctag
     2881 taattcttat atctgaagct gtcatcagca ctatcttgaa gtatacatgg caagctgaag
     2941 aaaaatggga tgaaccttgg tataaccaaa aaacagaaca tcaaagaaat agcagtaaga
     3001 ttctgagatt tatttcagac ttccttgctt ttttggttct ctacaatttc atcattccaa
     3061 tttcattata tgtgacagtc gaaatgcaga aatttcttgg atcgtttttt attggctggg
     3121 atcttgatct gtatcatgaa gaatcagatc agaaagctca agtcaatact tccgatctga
     3181 atgaagagct tggacaggta gagtacgtgt ttacagataa aactggtaca ctgacagaaa
     3241 atgagatgca gtttcgggaa tgttcaatta atggcatgaa ataccaagaa attaatggta
     3301 gacttgtacc cgaaggacca acaccagact cttcagaagg aaacttatct tatcttagta
     3361 gtttatccca tcttaacaac ttatcccatc ttacaaccag ttcctctttc agaaccagtc
     3421 ctgaaaatga aactgaacta attaaagaac atgatctctt ctttaaagca gtcagtctct
     3481 gtcacactgt acagattagc aatgttcaaa ctgactgcac tggtgatggt ccctggcaat
     3541 ccaacctggc accatcgcag ttggagtact atgcatcttc accagatgaa aaggctctag
     3601 tagaagctgc agcaaggatt ggtattgtgt ttattggcaa ttctgaagaa actatggagg
     3661 ttaaaactct tggaaaactg gaacggtaca aactgcttca tattctggaa tttgattcag
     3721 atcgtaggag aatgagtgta attgttcagg caccttcagg tgagaagtta ttatttgcta
     3781 aaggagctga gtcatcaatt ctccctaaat gtataggtgg agaaatagaa aaaaccagaa
     3841 ttcatgtaga tgaatttgct ttgaaagggc taagaactct gtgtatagca tatagaaaat
     3901 ttacatcaaa agagtatgag gaaatagata aacgcatatt tgaagccagg actgccttgc
     3961 agcagcggga agagaaattg gcagctgttt tccagttcat agagaaagac ctgatattac
     4021 ttggagccac agcagtagaa gacagactac aagataaagt tcgagaaact attgaagcat
     4081 tgagaatggc tggtatcaaa gtatgggtac ttactgggga taaacatgaa acagctgtta
     4141 gtgtgagttt atcatgtggc cattttcata gaaccatgaa catccttgaa cttataaacc
     4201 agaaatcaga cagcgagtgt gctgaacaat tgaggcagct tgccagaaga attacagagg
     4261 atcatgtgat tcagcatggg ctggtagtgg atgggaccag cctatctctt gcactcaggg
     4321 agcatgaaaa actatttatg gaagtttgca gaaattgttc agctgtatta tgctgtcgta
     4381 tggctccact gcagaaagca aaagtaataa gactaataaa aatatcacct gagaaaccta
     4441 taacattggc tgttggtgat ggtgctaatg acgtaagcat gatacaagaa gcccatgttg
     4501 gcataggaat catgggtaaa gaaggaagac aggctgcaag aaacagtgac tatgcaatag
     4561 ccagatttaa gttcctctcc aaattgcttt ttgttcatgg tcatttttat tatattagaa
     4621 tagctaccct tgtacagtat tttttttata agaatgtgtg ctttatcaca ccccagtttt
     4681 tatatcagtt ctactgtttg ttttctcagc aaacattgta tgacagcgtg tacctgactt
     4741 tatacaatat ttgttttact tccctaccta ttctgatata tagtcttttg gaacagcatg
     4801 tagaccctca tgtgttacaa aataagccca ccctttatcg agacattagt aaaaaccgcc
     4861 tcttaagtat taaaacattt ctttattgga ccatcctggg cttcagtcat gcctttattt
     4921 tcttttttgg atcctattta ctaataggga aagatacatc tctgcttgga aatggccaga
     4981 tgtttggaaa ctggacattt ggcactttgg tcttcacagt catggttatt acagtcacag
     5041 taaagatggc tctggaaact catttttgga cttggatcaa ccatctcgtt acctggggat
     5101 ctattatatt ttattttgta ttttccttgt tttatggagg gattctctgg ccatttttgg
     5161 gctcccagaa tatgtatttt gtgtttattc agctcctgtc aagtggttct gcttggtttg
     5221 ccataatcct catggttgtt acatgtctat ttcttgatat cataaagaag gtctttgacc
     5281 gacacctcca ccctacaagt actgaaaagg cacagcttac tgaaacaaat gcaggtatca
     5341 agtgcttgga ctccatgtgc tgtttcccgg aaggagaagc agcgtgtgca tctgttggaa
     5401 gaatgctgga acgagttata ggaagatgta gtccaaccca catcagcaga tcatggagtg
     5461 catcggatcc tttctatacc aacgacagga gcatcttgac tctctccaca atggactcat
     5521 ctacttgctc gagatatcta gacccagctt tcttgtacaa agtggttgat ctagagggcc
     5581 cgcggttcga actcgagtat ccatacgatg taccagacta cgcagagatc cactagttca
     5641 attccagctg agcgccggtc gctaccatta ccagttggtc tggtgtcaaa aataataata
     5701 accgggcagg ccatgtctgc ccgtatttcg cgtaaggaaa tccattatgt actatttaaa
     5761 ctcgagcggc cgccagcaca gtggtcgact gtggaatgtg tgtcagttag ggtgtggaaa
     5821 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5881 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5941 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6001 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6061 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6121 ttgcaaaaag cttcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg
     6181 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     6241 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     6301 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     6361 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     6421 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg aggatcgttt
     6481 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     6541 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     6601 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     6661 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     6721 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     6781 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6841 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6901 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6961 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     7021 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     7081 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     7141 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7201 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7261 cttgacgagt tcttctgagc gggactctgg ggttcgcatc gtcgacgata aaataaaaga
     7321 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     7381 tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt
     7441 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     7501 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     7561 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     7621 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     7681 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     7741 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     7801 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     7861 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     7921 ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta
     7981 agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc
     8041 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     8101 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8161 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8221 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8281 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8341 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8401 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8461 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8521 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8581 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8641 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8701 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8761 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8821 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8881 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg
     8941 cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     9001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     9061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     9121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     9181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     9241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     9301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     9361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     9421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9961 catttc
//
