LOCUS       sequence-307588-        8289 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses human VprBP N909 mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8289)
  AUTHORS   Nakagawa T, Lv L, Nakagawa M, Yu Y, Yu C, D'Alessio AC, Nakayama K, 
            Fan HY, Chen X, Xiong Y
  TITLE     CRL4(VprBP) E3 Ligase Promotes Monoubiquitylation and Chromatin 
            Binding of TET Dioxygenases.
  JOURNAL   Mol Cell. 2015 Jan 22;57(2):247-60. doi: 
            10.1016/j.molcel.2014.12.002. Epub 2014 Dec 31.
  PUBMED    25557551
REFERENCE   2  (bases 1 to 8289)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2014.12.002"; journalName: "Mol Cell"; date: 
            "2015-01-22- 22"; volume: "57"; issue: "2"; pages: "247-60"
COMMENT     Sequence Label: pcDNA3-HA-VprBP (N909)
FEATURES             Location/Qualifiers
     source          1..8289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             685..711
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    724..748
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(2016..2027)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(3550..3574)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(3608..3626)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3609..3626)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3646..3663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3652..3876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3922..4350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4009..4028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4219..4240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4359..4379)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4364..4693
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4544..4679
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4606..4625
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4760..5554
                     /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(4814..4833)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5424..5443
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5728..5849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5765..5784)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5819..5838
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5898..5914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5898..5914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5911..5933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5922..5938
                     /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(5946..5952,5953..5970,5971..5976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5991..6012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6129..6146)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6300..6888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6380..6399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7059..7850,7851..7919))
                     /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     7682..7701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7920..8024)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8099..8118)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac catgtaccca tacgacgtcc cagactacgc tgaattcgat
      721 atcacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc gactggatcc
      781 ggtaccgaat tcacaggcaa agccatgact acagtagtgg tacatgtgga ctccaaagct
      841 gagctcacta ccctgctgga gcagtgggaa aaggaacatg gcagtgggca ggacatggta
      901 cctatcctta ccaggatgtc tcaattgatt gaaaaagaaa ctgaagagta tcgtaaaggg
      961 gatccagacc catttgatga tcgacatcct ggtcgagctg atccagagtg tatgctgggc
     1021 cacttgctga gaatactctt caagaatgat gatttcatga atgcactggt gaatgcatat
     1081 gtgatgacaa gccgagagcc ccctttaaac actgcagctt gcagactcct attagacatc
     1141 atgccagggc tggaaactgc tgtcgtcttt caagaaaagg agggaattgt cgagaatctt
     1201 ttcaaatggg cccgagaggc cgatcaacca ttgaggacat attctactgg actgttagga
     1261 ggtgctatgg aaaatcaaga cattgctgcc aactatagag atgaaaattc acagctggtg
     1321 gcaatagtgc ttcgaagact gagggagcta cagctacagg aagtggcttt gcggcaggaa
     1381 aacaagcgtc ccagtccacg gaagctctct tctgaacccc ttttgcctct ggatgaggag
     1441 gctgtggata tggactatgg tgacatggct gtagatgtag tggatggaga ccaagaggaa
     1501 gcttctggag acatggagat ctcctttcat cttgattcag gccacaagac tagtagcaga
     1561 gtgaactcaa caaccaaacc tgaggatgga ggattaaaga aaaacaagtc agcaaaacag
     1621 ggtgacagag agaactttag gaaagccaag caaaagttgg gtttctcatc ttctgatcca
     1681 gatcgcatgt ttgttgagct gtctaatagc agttggtcag aaatgtctcc ctgggtgatt
     1741 ggcaccaatt atacccttta tcctatgact cctgctatcg agcagcgact cattctccaa
     1801 tatttgaccc ctctaggaga atatcaggag ctacttccca tattcatgca acttggatca
     1861 cgggagctga tgatgttcta tattgacctg aagcaaacta atgatgtcct gcttacattt
     1921 gaggcactaa agcacctagc atctctcctg ctccataaca aatttgccac agaatttgtt
     1981 gcgcatggtg gagtacagaa attactggaa atacctcgtc cttctatggc tgcaaccggt
     2041 gtgtctatgt gtttgtatta cctatcctac aatcaggatg ccatggaaag agtttgcatg
     2101 catccccaca atgttctgtc tgatgtggtg aactataccc tgtggttaat ggagtgttct
     2161 catgcttcag gatgctgcca tgctaccatg tttttttcaa tttgcttctc atttcgggcc
     2221 gtcttggagc tctttgaccg ctatgatggt cttcgtcgtc tggtgaactt gatcagtact
     2281 ttggagattc taaatttgga agatcagggt gcacttctga gtgatgatga aatatttgct
     2341 agccgccaaa ctgggaaaca tacctgcatg gccttgcgca aatactttga ggctcacctg
     2401 gccattaaat tggaacaagt gaagcagtca cttcagagga ctgagggtgg cattcttgtc
     2461 cacccacaac ccccgtacaa ggcatgctca tatactcatg aacagattgt ggaaatgatg
     2521 gaatttttga tagaatatgg cccagcgcag ctatattggg aaccagctga agttttcctc
     2581 aaactttctt gtgtgcaact cttgttgcag cttatttcta ttgcctgcaa ttggaagacc
     2641 tattatgcaa ggaatgacac tgtgcgcttt gctttggatg tcctggctat tcttactgtg
     2701 gtgccaaaaa tccagctcca gttggcagaa tcagtggacg tgttggatga ggctggatct
     2761 acagtctcta ctgtaggtat cagcattatt ttgggagtgg ctgagggtga gttcttcatc
     2821 catgatgctg aaattcagaa gtcagcactt cagattatca tcaattgtgt gtgtggccca
     2881 gataaccgaa tatccagtat tggtaaattt atctctggta ctcctcggag aaagctgcct
     2941 cagaacccta aaagcagtga gcacaccctg gccaagatgt ggaatgtggt tcagtccaac
     3001 aacggcatca aggtgctcct gtccttactg tccattaaga tgcccatcac agatgcagac
     3061 caaatccggg ccctggcctg caaagcccta gtgggcctgt ctcgcagtag cactgtccgg
     3121 cagatcatca gtaaactgcc ccttttcagc agctgccaga tccagcagct gatgaaggag
     3181 cctgtgctgc aggacaagcg cagtgaccat gtcaagttct gcaagtatgc tgctgaactc
     3241 attgaacggg tgtcaggaaa accacttctc attggcactg atgtttccct agcacgactg
     3301 cagaaagcag atgttgttgc ccagtcaagg atctccttcc ctgagaaaga gctgcttttg
     3361 ttgatacgaa accatcttat ttctaaaggg cttggagaaa cagcaaccgt gctgacaaaa
     3421 gaggctgacc tgcccatgac tgctgcctcc cattcttctg cctttacccc agtcactgct
     3481 gctgcttctc ctgtctctct accccgaacc cctcgtatcg ctaatggcat ttgaactcga
     3541 gatatctaga cccagctttc ttgtacaaag tggtgatatc ctcgagcatg catctagagg
     3601 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3901 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3961 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4021 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4081 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4141 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4201 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4261 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4321 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4441 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4501 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4561 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4621 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4681 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4741 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4801 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4861 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4921 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4981 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5041 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5101 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5161 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5221 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5281 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5341 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5401 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5461 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5521 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5581 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5641 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5701 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5761 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5821 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5881 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5941 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6001 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6061 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6121 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6181 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6241 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6301 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6361 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6421 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6481 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6541 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6601 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6661 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6721 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6781 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6841 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6901 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6961 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7021 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7081 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7141 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7201 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7261 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7321 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7381 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7441 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7501 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7561 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7621 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7681 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7741 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7801 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7861 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7921 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7981 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8041 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     8101 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     8161 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     8221 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     8281 atacgcgtt
//
