LOCUS       sequence-344454-        7930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector co-expressing dTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7930)
  TITLE     Oncofetal gene reactivation drives EZH2-mutant myeloid leukemia 
            pathogenesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7930)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRRL.PPT.SFFV.MCS.IRES.dTomato
FEATURES             Location/Qualifiers
     source          1..7930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..234
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             235..415
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    462..587
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1080..1313
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1498..1542
                     /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    1809..1926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1960..2366
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    2463..3015
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2607..2624)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2834..2853
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3027..3728
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3160..3178)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3428..3447
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3622..3645
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3963..4366
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             4456..4689
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4761..4882
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4798..4817)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4852..4871
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4922..5057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5077..5096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5078..5096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5106..5123)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5106..5122)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5115..5137)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5264..5719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5351..5370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5561..5582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5745..5849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5850..5918,5919..6710)
                     /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     complement(6068..6087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6881..7469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7370..7389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7623..7640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7757..7778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7793..7798,7799..7816,7817..7823)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7831..7847
                     /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)."
     primer_bind     7836..7858
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7855..7871
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7855..7871
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7890..7910
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7892..7910
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
      241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
      421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
      481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
      541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
      601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
      661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
      721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
      781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
      841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
      901 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
      961 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1021 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1081 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1141 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1201 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1261 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1321 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1381 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     1441 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     1501 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     1561 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     1621 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     1681 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     1741 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     1801 cggttaactt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     1861 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     1921 aattttatcg atcacgagac tagcctcgag ctagctgcag taacgccatt ttgcaaggca
     1981 tggaaaaata ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag
     2041 ctaacgttgg gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa
     2101 gaacagatgg tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga
     2161 tatggcccaa ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg
     2221 acctgaaatg accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt
     2281 cgcgcgcttc tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca
     2341 gtcctccgac agactgagtc ggccggtcga atcaagctta tcgataccgt cgacggatcc
     2401 accggtacta gtatactccg gacatatgac gcgtatgcat gagtaactga ggatccgccc
     2461 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     2521 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     2581 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     2641 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     2701 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     2761 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     2821 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     2881 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     2941 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     3001 aacacgatga taatatggcc acaaccatgg tgagcaaggg cgaggaggtc atcaaagagt
     3061 tcatgcgctt caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg
     3121 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     3181 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg
     3241 cgtacgtgaa gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct
     3301 tcaagtggga gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact
     3361 cctccctgca ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc
     3421 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt
     3481 acccccgcga cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg
     3541 gccactacct ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg
     3601 gctactacta cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg
     3661 tggaacagta cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc
     3721 tgtacaagta aagcggccgc gtcgacggat cccccgggct gcaggaattc gagcatctta
     3781 ccgccattta ttcccatatt tgttctgttt ttcttgattt gggtatacat ttaaatgtta
     3841 ataaaacaaa atggtggggc aatcatttac atttttaggg atatgtaatt actagttcag
     3901 gtgtattgcc acaagacaaa catgttaaga aactttcccg ttatttacgc tctgttcctg
     3961 ttaatcaacc tctggattac aaaatttgtg aaagattgac tgatattctt aactatgttg
     4021 ctccttttac gctgtgtgga tatgctgctt taatgcctct gtatcatgct attgcttccc
     4081 gtacggcttt cgttttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4141 tgtggcccgt tgtccgtcaa cgtggcgtgg tgtgctctgt gtttgctgac gcaaccccca
     4201 ctggctgggg cattgccacc acctgtcaac tcctttctgg gactttcgct ttccccctcc
     4261 cgatcgccac ggcagaactc atcgccgcct gccttgcccg ctgctggaca ggggctaggt
     4321 tgctgggcac tgataattcc gtggtgttgt cggggaagct gacgtccttt cgaattcgat
     4381 atcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4441 aagaaaaggg gggactggaa gggctacgta actcccaacg aagacaagat ctgctttttg
     4501 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     4561 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     4621 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     4681 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     4741 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     4801 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4861 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     4921 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4981 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5041 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     5101 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5161 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5221 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5281 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5341 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5401 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5461 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5521 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5581 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5641 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5701 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5761 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5821 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5881 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5941 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6001 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6061 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6121 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6181 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6241 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6301 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6361 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6421 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6481 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6541 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6601 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6661 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6721 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6781 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6841 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6901 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6961 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7021 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7081 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7141 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7201 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7261 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7321 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7381 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7441 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7501 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7561 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7621 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     7681 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     7741 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     7801 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     7861 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     7921 tggagctgca
//
