LOCUS       sequence-419577-        8727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8727)
  AUTHORS   Aoki K, Hyuga M, Tarumoto Y, Nishibuchi G, Ueda A, Ochi Y, Sugino S,
            Mikami T, Kobushi H, Kato I, Akahane K, Inukai T, Takaori-Kondo A, 
            Takita J, Ogawa S, Yusa K
  TITLE     Canonical BAF complex regulates the oncogenic program in human 
            T-cell acute lymphoblastic leukemia.
  JOURNAL   Blood. 2023 Nov 3:blood.2023020857. doi: 10.1182/blood.2023020857.
  PUBMED    37922452
REFERENCE   2  (bases 1 to 8727)
  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: "Blood. 2023
            Nov 3:blood.2023020857. doi: 10.1182/blood.2023020857."
COMMENT     Sequence Label: pKLV-EF1a-BFP-W
FEATURES             Location/Qualifiers
     source          1..8727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        109..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             630..810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    857..982
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1479..1712
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1897..1941
                     /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    2239..2356
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2827..4008
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3058..4000
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3957..3977
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4029..4730
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(4796..4816)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5214..5225)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5677..5694)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5683..5907
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5933..5953)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5933..5951)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5972..5988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5972..5988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5985..6007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5996..6012
                     /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(6020..6026,6027..6044,6045..6050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6065..6086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6203..6220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6454..6473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7133..7924,7925..7993))
                     /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     7756..7775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7994..8098)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8124..8579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8261..8282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8473..8492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8706..8727,1)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8720..8727,1..10)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     join(8721..8727,1..10)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgacta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      661 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      721 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      781 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      841 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      901 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      961 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1021 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1081 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1141 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1201 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1261 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1321 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1381 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1441 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1501 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1561 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1621 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1681 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1741 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1801 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1861 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1921 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1981 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2041 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2101 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2161 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2221 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2281 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2341 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
     2401 gggccctacg cgtgaggggc gcggaagccg gggcggagga ggagccaacc ccgggctggg
     2461 ctgagacccg cagaggaaga cgctctaggg atttgtcccg gactagcgag atggcaaggc
     2521 tgaggacggg aggctgattg agaggcgaag gtacacccta atctcaatac aacctttgga
     2581 gctaagccag caatggtaga gggaagattc tgcacgtccc ttccaggcgg cctccccgtc
     2641 accacccccc ccaacccgcc ccgaccggag ctgagagtaa ttcatacaaa aggactcgcc
     2701 cctgccttgg ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat
     2761 cgctgcgttc ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat
     2821 gcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     2881 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa agtggcgcgg ggtaaactgg
     2941 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     3001 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     3061 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     3121 tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     3181 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     3241 tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     3301 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     3361 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     3421 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     3481 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     3541 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     3601 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     3661 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     3721 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     3781 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     3841 ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca
     3901 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     3961 gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtggg aagcttggcg
     4021 cgcccaccat gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
     4081 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     4141 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     4201 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
     4261 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
     4321 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     4381 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
     4441 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
     4501 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat
     4561 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     4621 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     4681 tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattga gaattcgcgg
     4741 ccgctcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4801 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4861 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4921 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4981 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5041 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5101 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5161 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5221 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5281 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg caggtacctt
     5341 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5401 gactggaagg gctaattcac tcccaaagaa gtcaagatct gctttttgcc tgtactgggt
     5461 ctctctggtt agaccagagt ctctctggtt agaccagatc tgagcctggg agctctctgg
     5521 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     5581 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5641 gtggaaaatc tctagcagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5701 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5761 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5821 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagtc aatagcaggc
     5881 atgctgggga tgcggtgggc tctatgggcg gccgttaatt aagcttttgt tccctttagt
     5941 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6001 atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg
     6061 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6121 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6181 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6241 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6301 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6781 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8101 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     8161 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     8221 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     8281 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     8341 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     8401 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     8461 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     8521 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     8581 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     8641 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     8701 gttttcccag tcacgacgtt gtaaaac
//
