LOCUS       sequence-412780-        7833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CDC50B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7833)
  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 7833)
  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-CDC50B
FEATURES             Location/Qualifiers
     source          1..7833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1953..2156
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2107..2127
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2201..2220
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2201..2219
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2239..2256
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2257..2280
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(3386..3404)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3387..3404)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3424..3441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3430..3654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3700..4128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3787..3806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3997..4018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4142..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4384..4403
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4538..5332
                     /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(4592..4611)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5202..5221
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5506..5627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5543..5562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5597..5616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5676..5692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5676..5692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5689..5711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5700..5716
                     /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(5724..5730,5731..5748,5749..5754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5769..5790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5907..5924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6078..6666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6158..6177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6837..7628,7629..7697))
                     /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     7460..7479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7698..7802)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtactgat gaatccccta atgattttta tcaaaatcat taagttaagg
      541 tagatacaca tcttgtcata tgatcaaatg gtttcgccaa aaatcaataa tcagacaaca
      601 aaatgtgcga actcgatatt ttacacgact ctctttacca attctgcccc gaattacact
      661 taaaacgact caacagctta acgttggctt gccacgcctt acttgactgt aaaactctca
      721 ctcttaccga acttggccgt aacctgccaa ccaaagcgag aacaaaacat aacatcaaac
      781 gaatcgaccg attgttaggt aatcgtcacc tccacaaaga gcgactcgct gtataccgtt
      841 ggcatgctag ctttatctgt tcgggcaata cgatgcccat tgtacttgtt gactggtctg
      901 atatccgtga gcaaaaacgg cttatggtat tgcgagcttc agtcgcacta cacggtcgtt
      961 ctgttactct ttatgagaaa gcgttcccgc tttcagagca atgttcaaag aaagctcatg
     1021 accaatttct agccgacctt gcgagcattc taccgagtaa caccacaccg ctcattgtca
     1081 gtgatgctgg ctttaaagtg ccatggtata aatccgttga gaagctgggt tggtactggt
     1141 taagtcgagt aagaggaaaa gtacaatatg cagacctagg agcggaaaac tggaaaccta
     1201 tcagcaactt acatgatatg tcatctagtc actcaaagac tttaggctat aagaggctga
     1261 ctaaaagcaa tccaatctca tgccaaattc tattgtataa atctcgctct aaaggccgaa
     1321 aaaatcagcg ctcgacacgg actcattgtc accacccgtc acctaaaatc tactcagcgt
     1381 cggcaaagga gccatggatt ctagcaacta acttacctgt tgaaattcga acacccaaac
     1441 aacttgttaa tatctattcg aagcgaatgc agattgaaga aaccttccga gacttgaaaa
     1501 gtcctgccta cggactaggc ctacgccata gccgaacgag cagctcagag cgttttgata
     1561 tcatgctgct aatcgccctg atgcttcaac taacatgttg gcttgcgggc gttcatgctc
     1621 agaaacaagg ttgggacaag cacttccagg ctaacacagt cagaaatcga aacgtactct
     1681 caacagttcg cttaggcatg gaagttttgc ggcattctgg ctacacaata acaagggaag
     1741 acttactcgt ggctgcaacc ctactagctc aaaatttatt cacacatggt tacgctttgg
     1801 ggaaattatg aggggatctc tcagtgccaa gtacgccccc tattgacgtc aatgacggta
     1861 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     1921 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     1981 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     2041 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     2101 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     2161 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     2221 agacccaagc ttaccatgca ccaccaccac caccacgact acaaggacga cgatgacaag
     2281 cttggatctg ccgccatgac ctggagcgcc acggcccggg gcgcccacca gcccgacaac
     2341 accgccttca ctcagcagcg cctccccgcc tggcagccgc tgctgtcggc cagcatcgcg
     2401 ctgccgctct tcttctgcgc gggcctggcc ttcatcggcc tgggcctggg cctctactac
     2461 tcctccaacg gcatcaagga gctggagtac gactatacag gcgacccggg caccggtaac
     2521 tgctcggtgt gcgccgcggc tggccagggc cgggcgctgc cgcccccctg ctcgtgcgcc
     2581 tggtacttct cgctgcccga gctcttccag ggcccagtgt acctctacta cgagctgacc
     2641 aacttctacc agaacaaccg gcgctacggc gtgtcccgcg acgacgcgca gctgagcgga
     2701 ctccccagcg cgctgcgcca ccctgtcaac gagtgcgccc cctaccagcg cagcgcggcc
     2761 ggcctgccca tcgcgccctg cggcgccatc gccaacagcc tcttcaacga ctccttctcg
     2821 ctttggcacc agcgccagcc cggcgggccc tacgtcgagg tgccgctcga ccgctccggc
     2881 atcgcctggt ggaccgacta ccacgtcaag ttccgcaacc cgccgctggt caacggcagc
     2941 ctggcgttgg ccttccaggg cacggcgccc ccgcccaact ggcgccggcc agtctacgag
     3001 ctcagccccg accccaacaa caccggcttc atcaatcagg acttcgtggt gtggatgcgc
     3061 acggcggcgc tgcccacgtt ccgcaaactg tacgcgcgca tccgccaggg caactactcg
     3121 gccgggctgc cgcggggcgc ctaccgcgtc aacatcacct acaactaccc ggtgcgcgcg
     3181 ttcggcggcc acaagctcct catcttcagc agcatctcgt ggatgggtgg caagaacccc
     3241 ttcctgggca tcgcctacct ggtcgtcggc tccctctgca tcctcaccgg ctttgtcatg
     3301 ctggtcgtct acattcgcta ccaggaccag gacgacgacg acgaggagtg agcggccgct
     3361 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     3421 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3481 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3541 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3601 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     3661 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     3721 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3781 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3841 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3901 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3961 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4021 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4081 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     4501 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     4561 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4621 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4681 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     4741 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4801 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4861 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4921 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4981 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5041 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     5101 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5161 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5221 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     5281 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     5341 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     5401 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     5461 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     5521 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5581 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     5641 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5701 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     5761 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5821 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5881 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5941 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6001 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6061 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6121 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6181 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6241 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6301 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6361 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6421 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6481 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6541 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6601 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6661 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6721 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6781 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6841 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6901 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6961 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7021 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7081 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7141 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7201 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7261 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7321 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7381 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7441 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7501 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7561 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7621 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7681 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7741 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7801 cgcacatttc cccgaaaagt gccacctgac gtc
//
