LOCUS       sequence-239596-        7857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of shRNA targeting IKZF3 from a 
            miR30 embedded casette in the 3'UTR of ZsGreen-P2A-Puro insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  AUTHORS   Patil A, Manzano M, Gottwein E
  TITLE     CK1alpha and IRF4 are essential and independent effectors of 
            immunomodulatory drugs in primary effusion lymphoma.
  JOURNAL   Blood. 2018 Aug 9;132(6):577-586. doi: 10.1182/blood-2018-01-828418.
            Epub 2018 Jun 28.
  PUBMED    29954751
REFERENCE   2  (bases 1 to 7857)
  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: 
            "10.1182/blood-2018-01-828418"; journalName: "Blood"; date: 
            "2018-08-9- 9"; volume: "132"; issue: "6"; pages: "577-586"
COMMENT     Sequence Label: pZIP-hCMV-ZsGreen-P2A-Puro-IKZF3 sh1
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             389..646
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    693..818
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1315..1548
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1733..1777
                     /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    2040..2155
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2213..2516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2517..2720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2671..2691
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2717..2741
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2811..3503
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     CDS             3513..3569
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3570..4166
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     4063..4083
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     ncRNA           4187..4308
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4336..4350
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4378..4504
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     misc_feature    4538..5126
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4591..4611)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5009..5020)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5333..5566
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5657..5677)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5662..5991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5842..5977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5904..5923
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     rep_origin      complement(6080..6668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6160..6179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6839..7630,7631..7699))
                     /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     7462..7481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7700..7804)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac cagctagcga ggcgtggcct gggcgggact ggggagtggc
      421 gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta
      481 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      541 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      601 tagagatccc tccgaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
      661 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
      721 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
      781 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
      841 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
      901 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
      961 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1021 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1081 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1141 ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga
     1201 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1261 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1321 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1381 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1441 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1501 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1561 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1621 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1681 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1741 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1801 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1861 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1921 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1981 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcct
     2041 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     2101 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     2161 tttattacag ggacagcaga gatccagttt atcgataagc ttgggagttc cgcgttacat
     2221 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     2281 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     2341 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     2401 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     2461 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     2521 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     2581 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     2641 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     2701 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
     2761 gctgttttga cctccataga agacaccgac tctagctaga gtaccggtcc atggcccagt
     2821 ccaagcacgg cctgaccaag gagatgacca tgaagtaccg catggagggc tgcgtggacg
     2881 gccacaagtt cgtgatcacc ggcgagggca tcggctaccc cttcaagggc aagcaggcca
     2941 tcaacctgtg cgtggtggag ggcggcccct tgcccttcgc cgaggacatc ttgtccgccg
     3001 ccttcatgta cggcaaccgc gtgttcaccg agtaccccca ggacatcgtc gactacttca
     3061 agaactcctg ccccgccggc tacacctggg accgctcctt cctgttcgag gacggcgccg
     3121 tgtgcatctg caacgccgac atcaccgtga gcgtggagga gaactgcatg taccacgagt
     3181 ccaagttcta cggcgtgaac ttccccgccg acggccccgt gatgaagaag atgaccgaca
     3241 actgggagcc ctcctgcgag aagatcatcc ccgtgcccaa gcagggcatc ttgaagggcg
     3301 acgtgagcat gtacctgctg ctgaaggacg gtggccgctt gcgctgccag ttcgacaccg
     3361 tgtacaaggc caagtccgtg ccccgcaaga tgcccgactg gcacttcatc cagcacaagc
     3421 tgacccgcga ggaccgcagc gacgccaaga accagaagtg gcacctgacc gagcacgcca
     3481 tcgcctccgg ctccgccttg cccggatccg gcgcaacaaa cttctctctg ctgaaacaag
     3541 ccggagatgt cgaagagaat cctggaccga ccgagtacaa gcccacggtg cgcctcgcca
     3601 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     3661 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     3721 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     3781 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     3841 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     3901 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc
     3961 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     4021 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     4081 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     4141 gcatgacccg caagcccggt gcctgagcgg ccgcaatgga tcctgtttga atgaggcttc
     4201 agtactttac agaatcgttg cctgcacatc ttggaaacac ttgctgggat tacttcttca
     4261 ggttaaccca acagaaggct cgagaaggta tattgctgtt gacagtgagc gaaaatccct
     4321 tacagctatt caatagtgaa gccacagatg tattgaatag ctgtaaggga tttctgccta
     4381 ctgcctcgga cttcaagggg ctactttagg agcaattatc ttgtttacta aaactgaata
     4441 ccttgctatc tctttgatac atttttacaa agctgaatta aaatggtata aattaaatca
     4501 ctttagcttg cagatctacg cgtggcgcgc ctggaacaat caacctctgg attacaaaat
     4561 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4621 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4681 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4741 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4801 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4861 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4921 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4981 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5041 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5101 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa
     5161 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     5221 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     5281 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg
     5341 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     5401 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5461 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5521 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     5581 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct
     5641 tgacattgtt taaactggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6001 ggagcttgta tatccattac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7801 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cctcgcg
//
