LOCUS       sequence-420542-        8473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tagged DHB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8473)
  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 8473)
  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-DHB-mTurq2
FEATURES             Location/Qualifiers
     source          1..8473
                     /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             join(3108..3110,3111..3824)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3150..3171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3411..3430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3758..3779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3848..4436
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3901..3921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4319..4330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4643..5276
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5404..5420)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5404..5420)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5417..5439)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5428..5444
                     /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(5452..5458,5459..5476,5477..5482))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5497..5518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5635..5652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5806..6394)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5886..5905)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6565..7356,7357..7425))
                     /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     7188..7207
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7426..7530)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    7578..7712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7615..7634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7669..7688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(7880..8473,1..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 ccaccatgac aaatgatgtc acctggagcg
     2821 aggcctcttc gcctgatgag aggacactca cctttgctga aagatggcaa ttatcttcac
     2881 ctgatggagt agatacagat gatgatttac caaaatcgcg agcatccaaa agaacctgtg
     2941 gtgtgaatga tgatgaaagt ccaagcaaaa tttttatggt gggagaatct ccacaagtgt
     3001 cttccagact tcagaatttg agactgaata atttaattcc caggcaactt ttcaagccca
     3061 ccgataatca agaaactgga tccggggccc agggcagcgg cactagtgtg agcaagggcg
     3121 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3181 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3241 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgt
     3301 cctggggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     3361 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3421 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3481 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3541 actttagcga caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3601 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3661 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3721 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3781 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaacccggg cggggcgcgt
     3841 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3901 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3961 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4021 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4081 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4141 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4201 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     4261 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     4321 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     4381 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     4441 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     4501 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     4561 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4621 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc
     4681 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     4741 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     4801 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     4861 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     4921 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     4981 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     5041 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     5101 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5161 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5221 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     5281 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     5341 tgagaggcct tgacattgct agcgtttacc gtcgacctct agctagagct tggcgtaatc
     5401 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5461 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5521 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5581 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5641 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5701 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5761 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5821 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5881 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5941 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6001 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6061 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6121 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6181 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6241 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6301 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6361 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6421 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6481 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6541 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6601 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6661 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6721 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6781 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6841 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6901 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6961 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7021 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7081 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7141 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7201 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7261 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7321 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7381 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7441 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7501 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7561 cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     7681 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc
     7741 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac
     7801 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat
     7861 gtactacaaa cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt
     7921 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
     7981 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
     8041 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
     8101 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
     8161 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
     8221 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
     8281 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
     8341 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     8401 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     8461 tggtaactag aga
//
