LOCUS       sequence-380245-        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 shPRDM14-5
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     447..467
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             678..911
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    983..1104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1020..1039)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1074..1093
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1144..1279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1299..1318)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1300..1318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1328..1345)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1328..1344)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1337..1359)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1486..1941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1573..1592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1783..1804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1967..2071
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2072..2140,2141..2932)
                     /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(2290..2309)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3103..3691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3592..3611
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3845..3862
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3979..4000
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4015..4020,4021..4038,4039..4045)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4053..4069
                     /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     4058..4080
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4077..4093
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4077..4093
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4112..4132
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4114..4132
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4160..4386
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4387..4567
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4614..4739
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5232..5465
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5650..5694
                     /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        5992..6240
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5992..6012
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6163..6182
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6336..6453
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6502..7012
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6931..6949
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7034..7083,1..550)
                     /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(7034..7053)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
       61 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
      121 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
      181 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
      241 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
      301 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
      361 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
      421 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
      481 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
      541 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
      601 tgcatcggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
      661 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
      721 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      781 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      841 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      901 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
      961 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     1021 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1081 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     1141 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1201 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     1261 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
     1321 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     1381 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     1441 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     1501 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1561 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1621 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1681 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1741 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1801 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1861 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1921 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1981 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2041 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2101 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2161 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2221 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2281 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2341 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2401 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2461 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2521 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2581 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2641 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2701 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2761 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2821 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2881 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2941 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3001 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3061 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3121 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3181 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3241 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3301 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3361 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3421 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3481 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3541 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3601 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3661 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3721 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3781 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3841 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3901 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     3961 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     4021 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     4081 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     4141 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
     4201 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     4261 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     4321 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     4381 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4441 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4501 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4561 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     4621 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     4681 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     4741 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     4801 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     4861 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     4921 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     4981 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     5041 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     5101 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     5161 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     5221 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     5281 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     5341 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     5401 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     5461 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     5521 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     5581 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     5641 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     5701 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     5761 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     5821 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     5881 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     5941 tctcgacggt atcgatcacg agactagcct cgagcggccg cccccttcac cgagggccta
     6001 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
     6061 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     6121 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     6181 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     6241 gggtgggaaa tgtttctctc aatctcgaga ttgagagaaa catttcccac tttttaattc
     6301 tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     6361 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     6421 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     6481 ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca gccctgggtt
     6541 tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg
     6601 gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt
     6661 gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc
     6721 ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca
     6781 gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc
     6841 agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc
     6901 ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg
     6961 cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggatcc
     7021 accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     7081 ccc
//
