LOCUS       sequence-420507-        8711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Venus under CDC6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8711)
  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 8711)
  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-hCDC6p-Venus
FEATURES             Location/Qualifiers
     source          1..8711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     regulatory      3508..3517
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3514..3516,3517..3519,3520..4230)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3559..3580)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3820..3839)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4167..4188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4243..4263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4426..4650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4680..5268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4733..4753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5151..5162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5475..6108
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6236..6252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6236..6252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6249..6271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6260..6276
                     /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(6284..6290,6291..6308,6309..6314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6329..6350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6467..6484)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6638..7226)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6718..6737)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7397..8188,8189..8257))
                     /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     8020..8039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8258..8362)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8410..8544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8447..8466)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8501..8520
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataccggt caattcactg atgatcgtaa tattgttgct
     2221 tccctctttc taggctttat ggctctattt tgtttgttac tgaggggtaa aagataaatg
     2281 tttaccatca cctagaattg ggttctggcc tctaaaggaa cctgaggctt agatgaatta
     2341 ttggctttgg aagctggcct tcaaattact gcgctaattt atatttttca ttaaaaactc
     2401 agcttgcctc ttctatatag ctgtcttccc tggccctgaa accctagtgt ttcgccataa
     2461 aagattttaa aattaagggg tcataattcc ctccccatga tgtgtggatt aatggtaaga
     2521 agatgcacag aacataatat tcttaggttg aacgaaataa aagtaaagag ttggctctgt
     2581 ttctcacctt tgaagcacaa atcaagagat actatgatga agcatagttt ttctttatat
     2641 aggtgtgtag aactttacca taaaaatcac tagttcagcc atcaggagat ctggatccta
     2701 ggctcttcac tgtcaccaag atgctgtgac ctctaacctt gtatagaagt ttgctttgta
     2761 ctttgcgagg ttgagcatta gagaggtaag gaaagtgcct agcatcatac ctggcgcaca
     2821 gaacccaaaa cggtaggtat catgtagcag ttctgaaaat ctagcccatc aggatgatgc
     2881 aaatgggtac tttaggcagt gagaagggga accacatctt gacacttcca gtcgaaggaa
     2941 gagtgcgact gcgcggcagc aaagactacg cctcccagcg tgctttgcgg cgggccggcc
     3001 cgctttaccc agagtcgccc tgccgcaatc gcgcgtcttt ccaccgaggc cccggatgta
     3061 gattccctcc cccgttcagt ggtcgtggcc tcacagcgac tctaagactt ggggctctct
     3121 cattggctgt aactcttcca ctggattggt agcaaaaaaa gaggcggtgc ccaaggcgaa
     3181 aggctctgtg actacagcca atcagaatcg aggccgggct ttggcgggag gtgggaacgc
     3241 tgtggccatt cggatttggc gcgagcgcgg ctggagtttg ctgctgccgc tgtgcagttt
     3301 gttcaggggc ttgtggtggt gagtccgaga ggctgcgtgt gagagacgtg agaaggatcc
     3361 tgcactgagg aggtggaaag aagaggattg ctcgaggagg cctggggtct gtgaggcagc
     3421 ggagctgggt gaaggctgcg ggttccggcg aggcctgagg tgaagtgaag ggaaaagagc
     3481 tgagctttcg aaactagtaa ttcgagcgcc accatggtga gcaagggcga ggagctgttc
     3541 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     3601 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     3661 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
     3721 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     3781 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     3841 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3901 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3961 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     4021 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     4081 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     4141 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     4201 atcactctcg gcatggacga gctgtacaag ggaagtggtg gaccaaagaa gaagagaaag
     4261 gtaggtagtg gacatggctt cccgccggca gtggcagcac aggatgatgg cacgctgccc
     4321 atgtcttgtg cccaggagag cgggatggac cgtcaccctg cagcctgtgc ttctgctagg
     4381 atcaatgtgt aggaattctc tagacctagg ggatcctgat cacgactgtg ccttctagtt
     4441 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4501 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4561 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4621 ggcatgctgg ggatgcggtg ggctctatgg ctcgagcccg ggcggggcgc gtctggaaca
     4681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5041 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     5101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     5281 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     5341 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     5401 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5461 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     5521 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     5581 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     5641 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     5701 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     5761 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     5821 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     5881 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     5941 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6001 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6061 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6121 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     6181 cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6241 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6301 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6361 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6421 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6481 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6541 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6601 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6661 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6721 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6781 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6841 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6901 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6961 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7021 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7081 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7141 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7201 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7261 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7321 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7381 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7441 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7501 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7561 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7621 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7681 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7741 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7801 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7861 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7921 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7981 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8041 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8101 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8161 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8221 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8281 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8341 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     8401 cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     8461 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8521 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     8581 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     8641 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     8701 aacttagtag t
//
