LOCUS       sequence-380246-        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 shPRDM14-4
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     457..477
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             688..921
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    993..1114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1030..1049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1084..1103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1154..1289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1309..1328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1310..1328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1338..1355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1338..1354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1347..1369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1496..1951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1583..1602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1793..1814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1977..2081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2082..2150,2151..2942)
                     /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(2300..2319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3113..3701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3602..3621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3855..3872
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3989..4010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4025..4030,4031..4048,4049..4055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4063..4079
                     /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     4068..4090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4087..4103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4087..4103
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4122..4142
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4124..4142
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4170..4396
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4397..4577
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4624..4749
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5242..5475
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5660..5704
                     /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        6002..6250
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6002..6022
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6173..6192
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6346..6463
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6512..7022
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6941..6959
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7044..7083,1..560)
                     /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(7044..7063)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
       61 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
      121 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
      181 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
      241 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
      301 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
      361 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
      421 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
      481 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
      541 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
      601 cacgacccca tgcatcggta cctttaagac caatgactta caaggcagct gtagatctta
      661 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
      721 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      781 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      841 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      901 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
      961 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     1021 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1081 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     1141 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     1201 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     1261 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     1321 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     1381 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     1441 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     1501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1561 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1681 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1801 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     1861 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1921 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1981 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2041 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2101 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2161 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2221 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2281 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2341 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2401 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2461 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2521 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2581 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2641 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2701 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2761 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2821 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2881 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2941 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3781 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3841 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3901 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3961 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4021 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4081 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
     4141 gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg
     4201 caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg
     4261 ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct
     4321 gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct
     4381 agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4441 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4501 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     4561 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     4621 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     4681 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     4741 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     4801 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     4861 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     4921 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     4981 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     5041 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     5101 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     5161 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     5221 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     5281 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     5341 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     5401 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     5461 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     5521 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     5581 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     5641 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     5701 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     5761 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     5821 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     5881 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     5941 agtgaacgga tctcgacggt atcgatcacg agactagcct cgagcggccg cccccttcac
     6001 cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
     6061 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
     6121 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
     6181 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
     6241 acgaaacacc ggctggataa agactccctt caactcgagt tgaagggagt ctttatccag
     6301 tttttaattc tcgacctcga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     6361 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     6421 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     6481 cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt ttccaaggca
     6541 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
     6601 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
     6661 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
     6721 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
     6781 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
     6841 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
     6901 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
     6961 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
     7021 agggggatcc accggagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     7081 gcg
//
