LOCUS       sequence-380242-        7084 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive lentiviral expression of CALM2 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   Kim M, Ly SH, Xie Y, Duronio GN, Ford-Roshon D, Hwang JH, Sulahian 
            R, Rennhack JP, So J, Gjoerup O, Talamas JA, Grandclaudon M, Long 
            HW, Doench JG, Sethi NS, Giannakis M, Hahn WC
  TITLE     YAP1 and PRDM14 converge to promote cell survival and tumorigenesis.
  JOURNAL   Dev Cell. 2022 Jan 24;57(2):212-227.e8. doi: 
            10.1016/j.devcel.2021.12.006. Epub 2022 Jan 5.
  PUBMED    34990589
REFERENCE   2  (bases 1 to 7084)
  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.devcel.2021.12.006"; journalName: "Dev Cell"; date: 
            "2022-01-24- 24"; volume: "57"; issue: "2"; pages: "212-227.e8"
COMMENT     Sequence Label: pLKO.1 puro shCALM2-3
FEATURES             Location/Qualifiers
     source          1..7084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     466..486
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             697..930
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1002..1123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1039..1058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1093..1112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1163..1298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1318..1337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1319..1337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1347..1364)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1347..1363)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1356..1378)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1505..1960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1592..1611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1802..1823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1986..2090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2091..2159,2160..2951)
                     /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(2309..2328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3122..3710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3611..3630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3864..3881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3998..4019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4034..4039,4040..4057,4058..4064)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4072..4088
                     /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     4077..4099
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4096..4112
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4096..4112
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4131..4151
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4133..4151
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4179..4405
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4406..4586
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4633..4758
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5251..5484
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5669..5713
                     /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        6011..6259
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6011..6031
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6182..6201
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6356..6473
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6522..7032
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6951..6969
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7054..7084,1..569)
                     /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(7054..7073)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
       61 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
      121 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
      181 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
      241 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
      301 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
      361 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
      421 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
      481 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
      541 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
      601 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
      661 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
      721 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
      781 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      841 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      901 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt
      961 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     1021 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     1141 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     1201 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     1261 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     1321 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     1381 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     1441 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     1501 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1561 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1621 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1681 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1741 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1801 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1861 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     1921 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     1981 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2041 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2101 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2161 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2221 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2281 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2341 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2401 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2461 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2521 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2581 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2641 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2701 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2761 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2821 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2881 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2941 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3001 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3061 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3121 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3181 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3241 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3301 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3361 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3421 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3481 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3541 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3601 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3661 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3721 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3781 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3841 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3901 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3961 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4021 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4081 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     4141 ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt
     4201 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     4261 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     4321 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     4381 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     4441 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4501 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4561 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     4621 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     4681 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     4741 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     4801 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     4861 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     4921 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     4981 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     5041 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     5101 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     5161 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     5221 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     5281 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     5341 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     5401 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     5461 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     5521 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     5581 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     5641 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     5701 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     5761 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     5821 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     5881 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     5941 cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc
     6001 ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     6061 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     6121 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     6181 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     6241 gtggaaagga cgaaacaccg ggatggtcaa gtaaactatg aactcgagtt catagtttac
     6301 ttgaccatct ttttgaattc tcgacctcga gacaaatggc agtattcatc cacaatttta
     6361 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     6421 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     6481 attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt
     6541 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     6601 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     6661 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     6721 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     6781 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     6841 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     6901 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     6961 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     7021 ctctctcccc agggggatcc accggagctt accatgaccg agtacaagcc cacggtgcgc
     7081 ctcg
//
