LOCUS       sequence-380244-        7085 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 7085)
  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 7085)
  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 shCALM2-1
FEATURES             Location/Qualifiers
     source          1..7085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     460..480
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             691..924
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    996..1117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1033..1052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1087..1106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1157..1292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1312..1331)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1313..1331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1341..1358)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1341..1357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1350..1372)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1499..1954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1586..1605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1796..1817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1980..2084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2085..2153,2154..2945)
                     /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(2303..2322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3116..3704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3605..3624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3858..3875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3992..4013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4028..4033,4034..4051,4052..4058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4066..4082
                     /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     4071..4093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4090..4106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4090..4106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4125..4145
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4127..4145
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4173..4399
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4400..4580
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4627..4752
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5245..5478
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5663..5707
                     /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        6005..6253
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6005..6025
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6176..6195
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6351..6468
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6517..7027
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6946..6964
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7049..7085,1..563)
                     /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(7049..7068)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
       61 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
      121 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
      181 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
      241 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
      301 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
      361 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
      421 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
      481 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
      541 tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga
      601 gcgcacgacc ccatgcatcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
      661 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
      721 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
      781 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      841 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      901 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
      961 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     1021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     1141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     1201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     1261 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     1321 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     1381 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     1441 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     1501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1921 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2521 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3901 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3961 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4021 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4081 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     4141 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     4201 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     4261 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     4321 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     4381 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     4441 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4501 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4561 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     4621 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     4681 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     4741 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     4801 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     4861 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     4921 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     4981 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     5041 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
     5101 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     5161 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     5221 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     5281 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     5341 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     5401 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     5461 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     5521 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     5581 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     5641 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     5701 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     5761 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     5821 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     5881 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     5941 attagtgaac ggatctcgac ggtatcgatc acgagactag cctcgagcgg ccgccccctt
     6001 caccgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
     6061 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
     6121 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
     6181 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
     6241 aggacgaaac accggccagg tactcgtaca ctatttctcg agaaatagtg tacgagtacc
     6301 tggttttttg aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     6361 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     6421 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     6481 agggacagca gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca
     6541 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     6601 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     6661 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     6721 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     6781 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     6841 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     6901 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     6961 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     7021 tccccagggg gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     7081 caccc
//
