LOCUS       sequence-380248-        7083 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive lentiviral expression of PRDM14 shRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  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 7083)
  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 shPRDM14-3
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     450..470
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             681..914
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    986..1107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1023..1042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1077..1096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1147..1282
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1302..1321)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1303..1321)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1331..1348)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1331..1347)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1340..1362)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1489..1944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1576..1595)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1786..1807
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1970..2074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2075..2143,2144..2935)
                     /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(2293..2312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3106..3694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3595..3614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3848..3865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3982..4003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4018..4023,4024..4041,4042..4048)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4056..4072
                     /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     4061..4083
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4080..4096
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4080..4096
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4115..4135
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4117..4135
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4163..4389
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4390..4570
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4617..4742
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5235..5468
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5653..5697
                     /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        5995..6243
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5995..6015
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6166..6185
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6339..6456
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6505..7015
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6934..6952
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7037..7083,1..553)
                     /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(7037..7056)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
       61 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
      121 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
      181 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
      241 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
      301 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
      361 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
      421 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
      481 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
      541 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
      601 ccatgcatcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
      661 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
      721 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
      781 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
      841 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
      901 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
      961 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     1021 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1081 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     1141 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     1201 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     1261 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     1321 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1381 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1441 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     1501 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1561 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1621 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1681 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1741 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1801 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1861 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1921 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1981 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2041 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2101 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2161 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2221 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2281 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2341 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2401 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2461 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2521 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2581 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2641 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2701 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2761 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2821 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2881 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2941 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3001 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3721 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3781 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3841 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     3901 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     3961 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     4021 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     4081 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     4141 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     4201 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
     4261 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
     4321 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
     4381 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     4441 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     4501 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     4561 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     4621 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     4681 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     4741 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     4801 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     4861 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     4921 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     4981 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     5041 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     5101 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     5161 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     5221 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     5281 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     5341 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     5401 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     5461 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     5521 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     5581 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     5641 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     5701 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     5761 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     5821 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc
     5881 gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac
     5941 ggatctcgac ggtatcgatc acgagactag cctcgagcgg ccgccccctt caccgagggc
     6001 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
     6061 ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
     6121 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
     6181 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac
     6241 accggcggat ccacattctt catgttctcg agaacatgaa gaatgtggat ccgtttttaa
     6301 ttctcgacct cgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     6361 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     6421 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     6481 gatccacttt ggccgcggct cgagggggtt ggggttgcgc cttttccaag gcagccctgg
     6541 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     6601 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     6661 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     6721 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     6781 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     6841 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     6901 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     6961 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccaggggga
     7021 tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     7081 cgt
//
