LOCUS       sequence-365403-        7616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human XK in Expi293 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7616)
  AUTHORS   Adlakha J, Hong Z, Li P, Reinisch KM
  TITLE     Structural and biochemical insights into lipid transport by VPS13 
            proteins.
  JOURNAL   J Cell Biol. 2022 Jun 6;221(5):e202202030. doi: 
            10.1083/jcb.202202030. Epub 2022 Mar 31.
  PUBMED    35357422
REFERENCE   2  (bases 1 to 7616)
  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"; journalName: "J Cell 
            Biol."; date: "2022-06-6"; volume: "221(5):e202202030. doi"; pages: 
            " 10.1083/jcb.202202030. Epub 2022 Mar 31"
COMMENT     Sequence Label: pCMV10-3xFLAG-prs-XK
FEATURES             Location/Qualifiers
     source          1..7616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             997..1020
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    2375..2997
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2574..2593)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3021..3041)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3039..3336
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3390..4184
                     /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(3444..3463)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4054..4073
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4836..4970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4873..4892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4927..4946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4975..4994)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4976..4994)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5008..5024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5008..5024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5021..5043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5032..5048
                     /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(5056..5062,5063..5080,5081..5086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5101..5122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5239..5256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5410..5998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5490..5509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6169..6960,6961..7029))
                     /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     6792..6811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7030..7134)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7161..7589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7248..7267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7458..7479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctgg aagttctgtt ccaggggccc
     1021 cttgcggccg cgatgaaatt cccggcctcg gtgctggcgt ccgtgttcct gttcgtggcc
     1081 gagacaacgg cggcgctcag cctgagcagc acctaccgct cgggcgggga ccgcatgtgg
     1141 caggcgctga cgttgctttt ctcgctactg ccttgcgcgc tcgtgcagct cacgcttctc
     1201 ttcgtacacc gcgacctcag ccgcgaccgc ccgctcgtac tgctgctgca cctgctgcaa
     1261 cttgggcccc ttttcaggtg ttttgaagtc ttctgcatct actttcagtc aggcaacaat
     1321 gaagagcctt atgtcagtat caccaagaag aggcaaatgc caaaaaatgg cctctcagag
     1381 gagattgaga aggaggtggg ccaggcagaa ggcaaactaa tcacccaccg atcagcgttc
     1441 agccgggcgt cggtgatcca ggctttcttg ggctcagccc cccagctgac cctacagctg
     1501 tacataagtg tcatgcagca ggacgtcact gttggaagaa gtctcctcat gaccatatcc
     1561 ctgttgtcca ttgtgtatgg agccttgcgc tgcaacatcc tagccatcaa aatcaagtac
     1621 gatgagtatg aagtcaaagt gaagcctctg gcctatgtct gtatcttcct gtggaggagc
     1681 tttgagattg ccactcgagt tgtagtcctg gtcctcttta cctccgtcct gaagacctgg
     1741 gtggtggtta taatactcat caacttcttc agtttcttct tgtacccctg gatcctcttc
     1801 tggtgcagtg gttccccatt ccctgagaac atagagaagg ccctcagtag agtgggcacc
     1861 accattgtac tatgctttct aactttactc tatactggta tcaacatgtt ctgctggtct
     1921 gctgtacagc tgaaaattga cagccctgac ctcatcagca agtcccataa ttggtaccag
     1981 ctactggtgt attacatgat aagattcatc gagaatgcca tcctcctcct cctgtggtat
     2041 cttttcaaga ctgacatcta tatgtatgtg tgcgcacctc tgttggtcct gcagctgctc
     2101 attgggtact gcacagccat tctcttcatg cttgtattct atcagttctt ccacccttgc
     2161 aaaaagctct tttcttccag tgtttctgaa ggctttcaga ggtggctcag gtgtttttgc
     2221 tgggcctgca ggcagcaaaa accctgtgag ccgataggaa aggaagatct acagtcatcc
     2281 agagatagag atgagacacc ttctagcagt aaaacaagtc ctgagcctgg tcagttcttg
     2341 aatgctgaag atctctgctc tgcttaagga tcccgggtgg catccctgtg acccctcccc
     2401 agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa
     2461 attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg
     2521 ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg
     2581 ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt
     2641 tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag
     2701 gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt
     2761 ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca
     2821 ggcgtgaacc actgctccct tccctgtcct tctgatttta aaataactat accagcagga
     2881 ggacgtccag acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg
     2941 cttgcttggc actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg
     3001 tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3061 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3241 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3301 tgagctattc cagaagtagt gaggaggctt ttttggagga attgatcagc ttgggatctg
     3361 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3421 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3481 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3541 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3601 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3661 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3721 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3781 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3841 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3901 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3961 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4021 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4081 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4141 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4201 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4261 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4321 gcgcggggat ctcatgctgg agttcttcgc ccaccccggg ctcgatcccc tcgcgagttg
     4381 gttcagctgc tgcctgaggc tggacgacct cgcggagttc taccggcagt gcaaatccgt
     4441 cggcatccag gaaaccagca gcggctatcc gcgcatccat gcccccgaac tgcaggagtg
     4501 gggaggcacg atggccgctt tggtcgaccc ggacgggacg ctcctgcgcc tgatacagaa
     4561 cgaattgctt gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg
     4621 tggatggagc gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgccaagc
     4681 tggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccg atcataatca
     4741 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4801 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4861 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4921 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc aattccctat
     4981 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     5041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5221 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5281 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5341 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5821 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6841 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6901 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6961 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7021 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7081 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7141 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     7201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     7261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     7321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     7381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     7501 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     7561 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caattt
//
