LOCUS       sequence-380243-        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 shCALM2-2
FEATURES             Location/Qualifiers
     source          1..7085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     462..482
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             693..926
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    998..1119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1035..1054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1089..1108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1159..1294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1314..1333)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1315..1333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1343..1360)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1343..1359)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1352..1374)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1501..1956
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1588..1607)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1798..1819
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1982..2086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2087..2155,2156..2947)
                     /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(2305..2324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3118..3706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3607..3626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3860..3877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3994..4015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4030..4035,4036..4053,4054..4060)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4068..4084
                     /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     4073..4095
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4092..4108
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4092..4108
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4127..4147
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4129..4147
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4175..4401
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4402..4582
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4629..4754
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5247..5480
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5665..5709
                     /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        6007..6255
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6007..6027
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6178..6197
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6353..6470
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6519..7029
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6948..6966
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7051..7085,1..565)
                     /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(7051..7070)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
       61 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
      121 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
      181 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
      241 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
      301 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
      361 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
      421 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
      481 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
      541 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
      601 gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga
      661 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
      721 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
      781 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
      841 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
      901 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
      961 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     1021 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1081 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     1141 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1261 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     1321 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     1381 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     1441 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     1501 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1561 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1621 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1681 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1741 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1801 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1861 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1921 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     1981 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2041 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2101 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2161 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2221 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2281 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2341 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2401 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2461 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2521 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2581 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2641 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2701 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2761 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2821 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2881 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2941 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3001 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3061 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3181 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3241 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3301 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3361 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3421 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3481 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3541 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3601 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3661 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3721 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3781 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3841 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     3901 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     3961 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4021 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4081 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     4141 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     4201 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     4261 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     4321 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     4381 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     4441 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     4501 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     4561 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     4621 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     4681 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     4741 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     4801 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     4861 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     4921 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     4981 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     5041 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     5101 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     5161 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     5221 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     5281 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     5341 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     5401 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     5461 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     5521 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     5581 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     5641 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     5701 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     5761 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     5821 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     5881 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
     5941 cgattagtga acggatctcg acggtatcga tcacgagact agcctcgagc ggccgccccc
     6001 ttcaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     6061 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     6121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     6181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     6241 aaaggacgaa acaccgggaa gagtttgtac aaatgatgct cgagcatcat ttgtacaaac
     6301 tcttcttttt tgaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa
     6361 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     6421 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     6481 acagggacag cagagatcca ctttggccgc ggctcgaggg ggttggggtt gcgccttttc
     6541 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     6601 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     6661 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     6721 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     6781 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
     6841 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg
     6901 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc
     6961 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc
     7021 tctccccagg gggatccacc ggagcttacc atgaccgagt acaagcccac ggtgcgcctc
     7081 gccac
//
