LOCUS       sequence-267147-        7081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for enhancer reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  AUTHORS   Huang Y, Mouttet B, Warnatz HJ, Risch T, Rietmann F, Frommelt F, Ngo
            QA, Dobay MP, Marovca B, Jenni S, Tsai YC, Matzk S, Amstislavskiy V,
            Schrappe M, Stanulla M, Gstaiger M, Bornhauser B, Yaspo ML, Bourquin
            JP
  TITLE     The Leukemogenic TCF3-HLF Complex Rewires Enhancers Driving Cellular
            Identity and Self-Renewal Conferring EP300 Vulnerability.
  JOURNAL   Cancer Cell. 2019 Dec 9;36(6):630-644.e9. doi: 
            10.1016/j.ccell.2019.10.004. Epub 2019 Nov 14.
  PUBMED    31735627
REFERENCE   2  (bases 1 to 7081)
  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: 
            "10.1016/j.ccell.2019.10.004"; journalName: "Cancer Cell"; date: 
            "2019-12-9- 9"; volume: "36"; issue: "6"; pages: "630-644.e9"
COMMENT     Sequence Label: FpG5.BFP.Reporter
FEATURES             Location/Qualifiers
     source          1..7081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1837..1954
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             2221..2922
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    2938..3526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2991..3011)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3409..3420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3598..3831
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3909..4030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3946..3965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4000..4019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4070..4205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4225..4244)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4226..4244)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4254..4271)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4254..4270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4263..4285)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4412..4867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4709..4730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4893..4997
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4998..5066,5067..5858)
                     /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(5216..5235)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6029..6617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6518..6537
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6771..6788
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6905..6926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6941..6946,6947..6964,6965..6971)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6979..6995
                     /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     6984..7006
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7003..7019
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7003..7019
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7038..7058
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7040..7058
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     1741 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     1801 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     1861 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     1921 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     1981 tccagtttgg ttaattaagg atccgagctc ttacgcgtgc tagcccagca cccgggcgcg
     2041 gggcaggcga gtaggagggg gcgccggcta tatataccac tgctccggag ggctgggcgc
     2101 gcggggacta tcccgccacc gttgcgtccc tatttgctct cgctacttag gtctgtgcgc
     2161 agcactcacc gaactcacgg cccgcagctc gaactcacgc aagatccacc ggtcgccacc
     2221 atgagcgagc tgatcaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
     2281 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     2341 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
     2401 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
     2461 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     2521 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     2581 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     2641 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     2701 gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc
     2761 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     2821 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     2881 tactgcgacc tccctagcaa actggggcac aagcttaatt agacgcgtta agtcgacaat
     2941 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3001 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3061 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3121 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3181 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3241 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3301 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3361 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3421 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3481 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     3541 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     3601 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     3661 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     3721 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     3781 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     3841 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     3901 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3961 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4021 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     4081 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     4141 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     4201 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     4261 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     4321 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4381 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4441 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4501 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4561 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4621 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4681 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4741 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4801 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4861 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4921 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4981 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5041 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5101 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5161 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5221 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5281 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5341 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5401 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5461 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5521 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5581 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5641 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5701 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5761 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5821 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5881 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5941 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6001 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6061 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6121 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6481 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6541 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6601 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6661 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6721 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6781 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6841 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6901 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6961 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     7021 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     7081 c
//
