LOCUS       sequence-380252-        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 shPRDM14-1
FEATURES             Location/Qualifiers
     source          1..7083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     445..465
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             676..909
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    981..1102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1018..1037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1072..1091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1142..1277
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1297..1316)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1298..1316)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1326..1343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1326..1342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1335..1357)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1484..1939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1571..1590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1781..1802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1965..2069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2070..2138,2139..2930)
                     /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(2288..2307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3101..3689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3590..3609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3843..3860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3977..3998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4013..4018,4019..4036,4037..4043)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4051..4067
                     /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     4056..4078
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4075..4091
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4075..4091
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4110..4130
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4112..4130
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4158..4384
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4385..4565
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4612..4737
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5230..5463
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5648..5692
                     /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        5990..6238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5990..6010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6161..6180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    6334..6451
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6500..7010
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     6929..6947
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(7032..7083,1..548)
                     /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(7032..7051)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
       61 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
      121 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
      181 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
      241 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
      301 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
      361 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
      421 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
      481 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
      541 gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg
      601 catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
      661 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
      721 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      781 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
      841 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
      901 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
      961 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     1021 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1081 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     1141 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1201 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     1261 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     1321 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     1381 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     1441 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     1501 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1561 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1621 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1681 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1741 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1801 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1861 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1921 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2041 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2161 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2221 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2281 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2341 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2461 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2521 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2581 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2641 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2701 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2761 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2821 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2881 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2941 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3001 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3061 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3121 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3181 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3241 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3301 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3361 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3421 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3481 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3541 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3601 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3661 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3721 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3781 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3841 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3901 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3961 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     4021 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     4081 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     4141 tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     4201 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     4261 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     4321 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat
     4381 aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     4441 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     4501 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4561 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     4621 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     4681 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     4741 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     4801 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     4861 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     4921 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     4981 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     5041 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     5101 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     5161 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     5221 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     5281 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     5341 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     5401 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     5461 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     5521 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     5581 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     5641 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     5701 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     5761 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     5821 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg
     5881 aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc
     5941 tcgacggtat cgatcacgag actagcctcg agcggccgcc cccttcaccg agggcctatt
     6001 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     6061 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     6121 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     6181 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
     6241 cgtcctatgg acactacaga actcgagttc tgtagtgtcc ataggacgtt tttaattctc
     6301 gacctcgaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     6361 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     6421 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     6481 actttggccg cggctcgagg gggttggggt tgcgcctttt ccaaggcagc cctgggtttg
     6541 cgcagggacg cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt
     6601 ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat cttcgccgct acccttgtgg
     6661 gccccccggc gacgcttcct gctccgcccc taagtcggga aggttccttg cggttcgcgg
     6721 cgtgccggac gtgacaaacg gaagccgcac gtctcactag taccctcgca gacggacagc
     6781 gccagggagc aatggcagcg cgccgaccgc gatgggctgt ggccaatagc ggctgctcag
     6841 cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg
     6901 tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca
     6961 cgtcggcagt cggctccctc gttgaccgaa tcaccgacct ctctccccag ggggatccac
     7021 cggagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     7081 cca
//
