LOCUS       sequence-420566-        8827 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tagged H2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8827)
  AUTHORS   Konagaya Y, Rosenthal D, Ratnayeke N, Fan Y, Meyer T
  TITLE     An intermediate Rb-E2F activity state safeguards proliferation 
            commitment.
  JOURNAL   Nature. 2024 Jul;631(8020):424-431. doi: 10.1038/s41586-024-07554-2.
            Epub 2024 Jun 26.
  PUBMED    38926571
REFERENCE   2  (bases 1 to 8827)
  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: 
            "10.1038/s41586-024-07554-2"; journalName: "Nature"; date: 
            "2024-07"; volume: "631"; issue: "8020"; pages: "424-431"
COMMENT     Sequence Label: pLV-EF1a-H2B-miRFP670
FEATURES             Location/Qualifiers
     source          1..8827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    87..212
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    709..942
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1127..1171
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1434..1549
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        1627..1838
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          1839..2777
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2734..2754
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2799..3176
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             3231..4178
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     misc_feature    4202..4790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4255..4275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4673..4684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4997..5630
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5758..5774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5758..5774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5771..5793)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5782..5798
                     /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(5806..5812,5813..5830,5831..5836))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5851..5872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5989..6006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6160..6748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6240..6259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6919..7710,7711..7779))
                     /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     7542..7561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7780..7884)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7932..8066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7969..7988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8023..8042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(8234..8827,1..40)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
       61 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
      121 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
      181 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
      241 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
      301 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
      361 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
      421 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
      481 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
      541 cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg
      601 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
      661 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
      721 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
      781 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
      841 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
      901 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
      961 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1021 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     1081 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     1141 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     1201 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     1261 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     1321 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     1381 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gcctttaaaa
     1441 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     1501 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     1561 acagggacag cagagatcca gtttatcgat gaggcccttt cgtcttcact cgaggtgccc
     1621 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     1681 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     1741 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     1801 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     1861 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     1921 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     1981 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     2041 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     2101 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     2161 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     2221 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     2281 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     2341 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     2401 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     2461 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     2521 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     2581 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     2641 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     2701 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     2761 tttttcttcc atttcaggtg tcgtgaggat ccgccaccat gccagagcca gcgaagtctg
     2821 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
     2881 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
     2941 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaat tcgtttgtga
     3001 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct
     3061 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
     3121 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaagaaca
     3181 tggggcatgg caccggcagc accggcagcg gcagctcatc cggaactagt atggtagcag
     3241 gtcatgcctc tggcagcccc gcattcggga ccgcctctca ttcgaattgc gaacatgaag
     3301 agatccacct cgccggctcg atccagccgc atggcgcgct tctggtcgtc agcgaacatg
     3361 atcatcgcgt catccaggcc agcgccaacg ccgcggaatt tctgaatctc ggaagcgtac
     3421 tcggcgttcc gctcgccgag atcgacggcg atctgttgat caagatcctg ccgcatctcg
     3481 atcccaccgc cgaaggcatg ccggtcgcgg tgcgctgccg gatcggcaat ccctctacgg
     3541 agtactgcgg tctgatgcat cggcctccgg aaggcgggct gatcatcgaa ctcgaacgtg
     3601 ccggcccgtc gatcgatctg tcaggcacgc tggcgccggc gctggagcgg atccgcacgg
     3661 cgggttcact gcgcgcgctg tgcgatgaca ccgtgctgct gtttcagcag tgcaccggct
     3721 acgaccgggt gatggtgtat cgtttcgatg agcaaggcca cggcctggta ttctccgagt
     3781 gccatgtgcc tgggctcgaa tcctatttcg gcaaccgcta tccgtcgtcg actgtcccgc
     3841 agatggcgcg gcagctgtac gtgcggcagc gcgtccgcgt gctggtcgac gtcacctatc
     3901 agccggtgcc gctggagccg cggctgtcgc cgctgaccgg gcgcgatctc gacatgtcgg
     3961 gctgcttcct gcgctcgatg tcgccgtgcc atctgcagtt cctgaaggac atgggcgtgc
     4021 gcgccaccct ggcggtgtcg ctggtggtcg gcggcaagct gtggggcctg gttgtctgtc
     4081 accattatct gccgcgcttc atccgtttcg agctgcgggc gatctgcaaa cggctcgccg
     4141 aaaggatcgc gacgcggatc accgcgcttg agagctaacc cgggcggggc gcgtctggaa
     4201 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4261 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4321 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4381 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4441 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4501 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4561 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     4621 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     4681 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     4741 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     4801 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     4861 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     4921 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     4981 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     5041 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     5101 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     5161 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     5221 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     5281 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     5341 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     5401 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     5461 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     5521 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     5581 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     5641 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     5701 gccttgacat tgctagcgtt taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5761 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5821 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5881 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5941 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6001 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6061 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6901 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6961 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7021 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7081 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7141 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7201 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7261 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7321 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7381 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     7441 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7501 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7561 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7621 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7681 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7741 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7801 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7861 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7921 atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     7981 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8041 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
     8101 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
     8161 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
     8221 caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg
     8281 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg
     8341 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag
     8401 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg
     8461 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat
     8521 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac
     8581 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
     8641 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt
     8701 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     8761 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     8821 ctagaga
//
