LOCUS       sequence-442608-        8536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of FLAG-tagged human CAMK2D in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8536)
  AUTHORS   Chuah YH, Tay EXY, Grinchuk OV, Yoon J, Feng J, Kannan S, Robert M, 
            Jakhar R, Liang Y, Lee BWL, Wang LC, Lim YT, Zhao T, Sobota RM, Lu 
            G, Low BC, Crasta KC, Verma CS, Lin Z, Ong DST
  TITLE     CAMK2D serves as a molecular scaffold for RNF8-MAD2 complex to 
            induce mitotic checkpoint in glioma.
  JOURNAL   Cell Death Differ. 2023 Aug;30(8):1973-1987. doi: 
            10.1038/s41418-023-01192-3. Epub 2023 Jul 19.
  PUBMED    37468549
REFERENCE   2  (bases 1 to 8536)
  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: 
            "10.1038/s41418-023-01192-3"; journalName: "Cell Death Differ"; 
            date: "2023-08"; volume: "30"; issue: "8"; pages: "1973-1987"
COMMENT     Sequence Label: plenti-FLAG-CAMK2D
FEATURES             Location/Qualifiers
     source          1..8536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2..119
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(417..461)
                     /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    646..879
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1376..1501
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1548..1728
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1746..1944)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1946..2325
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2496..2515
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2590..2694
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2695..3354
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2761..2780)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      3431..4019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3920..3939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             4081..4261
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     4770..4786
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4771..4787
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4790..5378
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4896..4907
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5305..5325
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    5434..5467
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5524..6123)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5607..5627)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6104..6123
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6124..6180)
                     /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             complement(6199..6222)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(6241..6270)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(7827..7846)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7828..7846)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7872..8075)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        8076..8455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta
       61 ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat
      121 tgtggatgaa tactgccatt tgtctgcaga attggcgcac gcagtgccga tccgttcact
      181 aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg
      241 tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta
      301 actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact
      361 atcattatga ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa
      421 cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat
      481 tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc
      541 caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc
      601 acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc
      661 tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg
      721 agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact
      781 ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg
      841 cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct
      901 ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact
      961 actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg
     1021 aagatcagcg gccggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta
     1081 tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg
     1141 ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta
     1201 gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga
     1261 atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc
     1321 ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc
     1381 tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca
     1441 gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga
     1501 gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt
     1561 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
     1621 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
     1681 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccag tacaggcaaa
     1741 acgcgctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     1801 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     1861 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     1921 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     1981 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     2041 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     2101 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     2161 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     2221 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     2281 accccgtaat tgattactat taataactag tcaataatca atgtcaacgc gatatctggc
     2341 ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca tgcaattgtc
     2401 ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag cgacctccaa
     2461 cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga ttgtactgag
     2521 agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg cacttttcgg
     2581 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2641 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatggag
     2701 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     2761 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     2821 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     2881 cttgcccgcc tgatgaatgc tcatccggag ttccgtatgg caatgaaaga cggtgagctg
     2941 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     3001 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     3061 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     3121 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     3181 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     3241 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     3301 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatcatga
     3361 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3421 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3481 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3541 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3901 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3961 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4021 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgc
     4081 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     4141 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     4201 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     4261 cagtacagtc cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg ctgtcaaacc
     4321 tccactctaa tacttctctc tccgggtcat ccatcccatg caggctcaca gggtgtaaca
     4381 agcgggtgtt ctctccttca ttggcttctt ctaccttctc ttgctcaact ggtactagct
     4441 tgtagcacca tccaaaggtc agtggatatc tgatccctgg ccctggtgtg tagttctgcc
     4501 aatcagggaa gtagccttgt gtgtggtaga tccacagatc aaggatatct tgtcttcgtt
     4561 gggagtgaat tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag
     4621 ctgccttgta agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct
     4681 cctcctcttg tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg
     4741 tgattgctcc atgtttttct aggtctcgat cgaggtcgac ggtatcgatg cggggaggcg
     4801 gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga
     4861 gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag
     4921 aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat
     4981 gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag
     5041 cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc
     5101 ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac
     5161 acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc
     5221 aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac
     5281 aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca
     5341 gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat
     5401 tgtacctagt ggaaccggaa cccttaaaca tgtataactt cgtataatgt atgctatacg
     5461 aagttattag gtccctcgac gaatcctact tgtacagacc gcggccggcc gtttgggtgt
     5521 acatcaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac
     5581 gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt
     5641 ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc
     5701 gctgcccaga cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc
     5761 gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg
     5821 ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac
     5881 gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc
     5941 gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg
     6001 gcgatccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt
     6061 gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt
     6121 cataggtcca gggttctcct ccacgtctcc agcctgcttc agcaggctga agttagtagc
     6181 tccgcttcct acccaaacct tatcgtcgtc atccttgtaa tccaggatat catttgctgc
     6241 cagatcctct tctgagatga gtttctgctc gaggatgttt tgccacaaag aggtgcctcc
     6301 tgagaagttt tctttcccat ttggaataca gggtggcttg atgggtactg ttggtgaccc
     6361 cgagcgatga aaatgaacat tctgccactt tccatcccgg cggtgccaca cacgagtctc
     6421 ttctgactgc attgtctttg gcattccact gccatccatg tactgtgtga gcctaatata
     6481 tgctatgcag gcggcatcat cccctaccag atgtacatga gggtttagaa taatagtgtg
     6541 gattggttta ttgcttttgg acaaagcatt ttcaaagtag aatcggtgaa aatccatccc
     6601 ttccactaaa ttacccaaag cttcaggttc aaaagcagta aggcctgggt cacagatttt
     6661 tgtgtaggct tcaaagtccc cattgttgat agcttcgatc agttgttcag tgactttgat
     6721 aatctcttgc tttcgtgctt tcacatcttc atcctcaatt gttgtatttg aactctcagt
     6781 tgactccttt actccatctg gtttcttcaa caaactcttg gctgctgaga aattccttgt
     6841 agccagcata gttgtcaaga tggcaccctt tagttttctt ctagcattaa atttcttcaa
     6901 gcagtctaca gtctcctgtc tgtgcatcat ggaagcaaca gtagaacgtt gacagatcca
     6961 tgggtgcttc agtgcctctg aggctgtgat gcgtttggca gggttgatag taagcatttt
     7021 attgatgagg tctttggctt caggagtcac cgtgtcccat tctggtgatg gaaaatcata
     7081 agctccagcc ttgatctgct gatagagtct gtgttggtct tcatcccaga agggtggata
     7141 ccccacaagt agaatataga gaatgacacc acatgcccac atatccactg gctttccata
     7201 aggatcttta cgtaaaactt ctggagaaag atatccaggt gtgccagcaa aaccaaacca
     7261 cgcctgctgg tccccttgaa cttctatggc taagccaaag tctgccaatt tcacagctgc
     7321 tcccttggat ttgctagcta aaagcaaatt ctcaggcttc aggtccctgt gaactatgcc
     7381 atttaggtga caatgattaa cactttctag aatctgctgt atacaatgac tggcatcagc
     7441 ttcactgtag tattctcttg ccactatgtc ttcaaacagt tcacctccag taactaaatc
     7501 aaacaccaag tagtgaaagc cctcttctga tatgctatca tgaagtcgca caatattagg
     7561 gtgcttcaaa agacggcaga ttctagcttc tctttctagt ttctgatgat ccctagcaga
     7621 aagctttttg gtgttgataa ttttggcagc atattcttgt ccagtaggaa ttttcataca
     7681 tcttctcacc actgagaatg ccccctttcc aagctcctcg aaaagctgat actcgtccgt
     7741 gaacctggtg caggttgtgg tcgaagccat gcgatcgcgg cggcagatct cctcggtacc
     7801 ggatccagtc gacgaattcc cggccgccct atagtgagtc gtattacaaa attctgacgg
     7861 ttcactaaac gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     7921 gtcaacgggg cggggttatt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     7981 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     8041 ccacgcccat tggtgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     8101 tagatgtact gccaagtagg aaagtcccgt aaggtcatgt actgggcata atgccaggcg
     8161 ggccatttac cgtcattgac gtcaataggg ggcggacttg gcatatgata cacttgatgt
     8221 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     8281 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     8341 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     8401 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacat
     8461 ggcggtcata ttggacatgt ctagagcggg cccggtacta accaaactgg atctctgctg
     8521 tccctgtaat aaaccc
//
