LOCUS       sequence-412779-        6525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CDC50A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6525)
  AUTHORS   Takatsu H, Baba K, Shima T, Umino H, Kato U, Umeda M, Nakayama K, 
            Shin HW
  TITLE     ATP9B, a P4-ATPase (a putative aminophospholipid translocase), 
            localizes to the trans-Golgi network in a CDC50 protein-independent 
            manner.
  JOURNAL   J Biol Chem. 2011 Nov 4;286(44):38159-38167. doi: 
            10.1074/jbc.M111.281006. Epub 2011 Sep 13.
  PUBMED    21914794
REFERENCE   2  (bases 1 to 6525)
  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.M111.281006"; journalName: "J Biol Chem"; date: 
            "2011-11-4- 4"; volume: "286"; issue: "44"; pages: "38159-38167"
COMMENT     Sequence Label: pcDNA3/HF-CDC50A
FEATURES             Location/Qualifiers
     source          1..6525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     465..484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(703..807)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(882..901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1073..1452
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1453..1656
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1607..1627
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1701..1720
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1701..1719
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1739..1756
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1757..1780
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2916..2934)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2917..2934)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2954..2971)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2960..3184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3230..3658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3317..3336)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3527..3548
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3667..3687)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3672..4001
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3852..3987
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3914..3933
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4068..4862
                     /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(4122..4141)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4732..4751
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5036..5157
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5073..5092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5127..5146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5206..5222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5206..5222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5219..5241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5230..5246
                     /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(5254..5260,5261..5278,5279..5284))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5299..5320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5437..5454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5608..6196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5688..5707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6367..6525,1..633,634..702))
                     /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"
ORIGIN
        1 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
       61 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      121 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      181 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      241 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      301 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      361 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      421 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      481 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
      541 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
      601 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
      661 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
      721 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      781 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
      841 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
      901 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
      961 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     1021 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     1081 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1141 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1201 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1261 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1321 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1381 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1441 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     1501 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1561 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1621 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     1681 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttaccatgca
     1741 ccaccaccac caccacgact acaaggacga cgatgacaag cttggatctg ccaccatggc
     1801 gatgaactat aacgcgaagg atgaagtgga cggtgggccc ccgtgtgctc cggggggcac
     1861 cgcgaagact cggagaccgg ataacacggc cttcaaacag caacggctgc cagcttggca
     1921 gcccatcctt acggctggca cggtgctacc tattttcttc atcatcggtc tcatcttcat
     1981 tcccatcggc attggcatct ttgtcacctc caacaacatc cgcgagatcg agattgatta
     2041 taccggaaca gagccttcca gtccctgtaa taaatgttta tctccggatg tgacaccttg
     2101 cttttgtacc attaacttca cactggaaaa gtcatttgag ggcaacgtgt ttatgtatta
     2161 tggactgtct aatttctatc aaaaccatcg tcgttacgtg aaatctcgag atgatagtca
     2221 actaaatgga gattctagtg ctttgcttaa tcccagtaag gaatgtgaac cttatcgaag
     2281 aaatgaagac aaaccaattg ctccttgtgg agctattgcc aacagcatgt ttaatgatac
     2341 attagaattg tttctcattg gcaatgattc ttatcctata cctatcgctt tgaaaaagaa
     2401 aggtattgct tggtggacag ataaaaatgt gaaattcaga aatccccctg gaggagacaa
     2461 cctggaagaa cgatttaaag gtacaacaaa gcctgtgaac tggcttaaac cagtttacat
     2521 gctggattct gacccagata ataatggatt cataaatgag gattttattg tttggatgcg
     2581 tactgcagca ttacctactt ttcgcaagtt gtatcgtctt atagaaagga aaagtgattt
     2641 acatccaaca ttaccagctg gccgatactc tttgaatgtc acatacaatt accctgtaca
     2701 ttattttgat ggacgaaaac ggatgatctt gagcactatt tcatggatgg gaggaaaaaa
     2761 tccatttttg gggattgctt acatcgctgt tggatccatc tccttccttc tgggagttgt
     2821 actgctagta attaatcata aatatagaaa cagtagtaat acagctgaca ttaccattta
     2881 agcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     2941 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3001 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3061 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3121 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3181 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3241 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3301 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3361 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3421 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3481 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3541 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3601 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3661 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3721 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3781 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3841 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3901 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3961 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4021 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4081 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4141 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4201 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4261 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4321 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4381 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4441 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4501 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4561 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4621 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4681 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4741 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4801 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4861 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4921 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4981 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5041 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5101 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5161 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5221 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5281 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5341 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5401 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5461 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5521 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5581 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5641 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5701 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5761 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5821 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5881 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5941 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6001 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6061 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6121 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6181 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6241 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6301 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6361 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6421 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6481 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggc
//
