LOCUS       sequence-239599-        7954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of Flag tagged CK1alpha 
            G40N-P2A-Blast casette from a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7954)
  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 7954)
  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: pLC-Flag-CSNK1A1 G40N-P2A-Blast
FEATURES             Location/Qualifiers
     source          1..7954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..396
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(416..449)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    505..1093
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(558..578)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(976..987)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1096..1112)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1097..1113)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1622..1802
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(1864..2452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1944..1963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2623..3414,3415..3483))
                     /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     3246..3265
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3484..3588)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3663..3682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3854..4233
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4235..4433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             4451..4631
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4678..4803
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5300..5533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5718..5762
                     /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    6060..6177
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        6302..6605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6606..6809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      6848..6857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6857..6880
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7898..7954
                     /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"
ORIGIN
        1 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
       61 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
      121 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
      181 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
      241 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
      301 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
      361 gaattgctgc cctctggtta tgtgtgggag ggctaagaat tcgtcgaggg acctaataac
      421 ttcgtatagc atacattata cgaagttata catgtttaag ggttccggtt ccactaggta
      481 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
      541 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
      601 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
      661 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
      721 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
      781 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
      841 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
      901 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
      961 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     1021 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     1081 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     1141 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     1201 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     1261 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     1321 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     1381 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     1441 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     1501 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     1561 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     1621 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     1681 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     1741 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     1801 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2641 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2701 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2761 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2821 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2881 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2941 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3001 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3061 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3121 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3181 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3241 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3301 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3361 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3421 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3481 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3541 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3601 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     3661 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     3721 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     3781 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     3841 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     3901 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     3961 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     4021 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     4081 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     4141 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     4201 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     4261 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4321 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     4381 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt
     4441 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4501 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4561 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4621 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     4681 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     4741 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     4801 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     4861 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     4921 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     4981 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5041 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5101 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccggccg ctgatcttca
     5161 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     5221 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     5281 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     5341 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     5401 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     5461 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     5521 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     5581 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     5641 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     5701 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     5761 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     5821 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     5881 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     5941 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     6001 tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt catccacaat
     6061 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     6121 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     6181 gtttattaca gggacagcag agatccagtt tggttagtac cgggcccgag agttaattaa
     6241 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6301 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     6361 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     6421 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6481 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6541 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6601 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6661 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6721 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6781 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     6841 accggtcgcc accatggatt acaaagacga tgacgataag gcgagtagca gcggctccaa
     6901 ggctgaattc attgtcggag ggaaatataa actggtacgg aagatcgggt ctggctcctt
     6961 cggggacatc tatttggcga tcaacatcac caacaacgag gaagtggcag tgaagctaga
     7021 atctcagaag gccaggcatc cccagttgct gtacgagagc aagctctata agattcttca
     7081 aggtggggtt ggcatccccc acatacggtg gtatggtcag gaaaaagact acaatgtact
     7141 agtcatggat cttctgggac ctagcctcga agacctcttc aatttctgtt caagaaggtt
     7201 cacaatgaaa actgtactta tgttagctga ccagatgatc agtagaattg aatatgtgca
     7261 tacaaagaat tttatacaca gagacattaa accagataac ttcctaatgg gtattgggcg
     7321 tcactgtaat aagttattcc ttattgattt tggtttggcc aaaaagtaca gagacaacag
     7381 gacaaggcaa cacataccat acagagaaga taaaaacctc actggcactg cccgatatgc
     7441 tagcatcaat gcacatcttg gtattgagca gagtcgccga gatgacatgg aatcattagg
     7501 atatgttttg atgtatttta atagaaccag cctgccatgg caagggctaa aggctgcaac
     7561 aaagaaacaa aaatatgaaa agattagtga aaagaagatg tccacgcctg ttgaagtttt
     7621 atgtaagggg tttcctgcag aatttgcgat gtacttaaac tattgtcgtg ggctacgctt
     7681 tgaggaagcc ccagattaca tgtatctgag gcagctattc cgcattcttt tcaggaccct
     7741 gaaccatcaa tatgactaca catttgattg gacaatgtta aagcagaaag cagcacagca
     7801 ggcagcctct tccagtgggc agggtcagca ggcccaaacc cccacaggca agcaaactga
     7861 caaaaccaag agtaacatga aaggtttcgg atccggcgca acaaacttct ctctgctgaa
     7921 acaagccgga gatgtcgaag agaatcctgg accg
//
