LOCUS       sequence-194636-        7607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentiviral vector expressing IKZF1 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7607)
  AUTHORS   Lu G, Middleton RE, Sun H, Naniong M, Ott CJ, Mitsiades CS, Wong KK,
            Bradner JE, Kaelin WG Jr
  TITLE     The myeloma drug lenalidomide promotes the cereblon-dependent 
            destruction of Ikaros proteins.
  JOURNAL   Science. 2014 Jan 17;343(6168):305-9. doi: 10.1126/science.1244917. 
            Epub 2013 Nov 29.
  PUBMED    24292623
REFERENCE   2  (bases 1 to 7607)
  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: 
            "10.1126/science.1244917"; journalName: "Science"; date: 
            "2014-01-17- 17"; volume: "343"; issue: "6168"; pages: "305-9"
COMMENT     Sequence Label: pLKO-RFP-IKZF1-sh2
FEATURES             Location/Qualifiers
     source          1..7607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    270..291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(306..311,312..329,330..336)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    344..360
                     /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     349..371
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     368..384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     368..384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     403..423
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        405..423
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        451..677
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             678..858
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    905..1030
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1523..1756
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1941..1985
                     /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"
     promoter        2163..2411
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2163..2183
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2334..2353
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2508..2625
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2674..3184
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3103..3121
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3199..3894
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     misc_feature    3910..4498
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3963..3983)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4381..4392)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4570..4803
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4881..5002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4918..4937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4972..4991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5042..5177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5197..5216)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5198..5216)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5226..5243)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5226..5242)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5235..5257)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5384..5839
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5471..5490)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5681..5702
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5865..5969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5970..6038,6039..6830)
                     /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(6188..6207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7001..7589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7490..7509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
       61 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      301 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      361 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      421 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
      481 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
      541 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
      601 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
      661 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
      721 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
      781 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
      841 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
      901 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
      961 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     1021 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     1081 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     1141 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     1201 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     1261 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     1321 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     1381 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     1441 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     1501 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     1561 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     1621 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     1681 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     1741 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     1801 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     1861 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     1921 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     1981 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     2041 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     2101 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccca
     2161 gagagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     2221 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     2281 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     2341 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     2401 gacgaggtac cggctacgag aaggagaacg aaatctcgag atttcgttct ccttctcgta
     2461 gtttttgaat tctagatctt gagacaaatg gcagtattca tccacaattt taaaagaaaa
     2521 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2581 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2641 gacagcagag atccactttg gcgccggctc gagggggccc gggttgcgcc ttttccaagg
     2701 cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg
     2761 cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc
     2821 cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc
     2881 cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct
     2941 cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa
     3001 tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc
     3061 ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa
     3121 gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc
     3181 ccagggggat ccgctagcat gagcgagctg atcaaggaga acatgcacat gaagctgtac
     3241 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
     3301 tacgagggca cccagaccat gaagatcaag gtggtcgagg gcggccctct ccccttcgcc
     3361 ttcgacatcc tggctaccag cttcatgtac ggcagcaaag ccttcatcaa ccacacccag
     3421 ggcatccccg acttctttaa gcagtccttc cctgagggct tcacatggga gagaatcacc
     3481 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcttcca gaacggctgc
     3541 atcatctaca acgtcaagat caacggggtg aacttcccat ccaacggccc tgtgatgcag
     3601 aagaaaacac gcggctggga ggccaacacc gagatgctgt accccgctga cggcggcctg
     3661 agaggccaca gccagatggc cctgaagctc gtgggcgggg gctacctgca ctgctccttc
     3721 aagaccacat acagatccaa gaaacccgct aagaacctca agatgcccgg cttccacttc
     3781 gtggaccaca gactggaaag aatcaaggag gccgacaaag agacctacgt cgagcagcac
     3841 gagatggctg tggccaagta ctgcgacctc cctagcaaac tggggcacag ataaacgcgt
     3901 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3961 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4021 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4081 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4141 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4201 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4261 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     4321 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4381 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4441 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     4501 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     4561 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     4621 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     4681 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     4741 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     4801 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     4861 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     4921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4981 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     5041 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5101 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5161 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     5221 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5281 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5341 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5401 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5461 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5521 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5581 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5641 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5701 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5761 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5821 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5881 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5941 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6001 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6061 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6121 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6181 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6241 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6301 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6361 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6421 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6481 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6541 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6601 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6661 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6721 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6781 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     6841 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6901 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7141 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7381 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggcc
//
