LOCUS       sequence-380325-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive retroviral expression of PRDM14.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  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 7479)
  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: pMSCV puro PRDM14 Delta-Zinc finger domain
FEATURES             Location/Qualifiers
     source          1..7479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1260..1282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1382..1401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1595..1617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1745..1764
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             1964..2480
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2544..2885
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2952..3368
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3298..3320
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3336..3352
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      3387..3396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3396..3419
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        4630..5117
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4665..4688)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5050..5069
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5138..5737
                     /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(5138..5157)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5634..5654
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5837..6351
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6513..6529
                     /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)."
     promoter        complement(join(6537..6543,6544..6561,6562..6567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6582..6603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6720..6737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6891..7479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6971..6990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1261 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1321 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1441 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     1501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1621 gcgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     1681 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     1741 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     1801 ccacgatgcg tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg
     1861 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     1921 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     1981 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa atacataact
     2041 gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata tgggccaaac
     2101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     2161 atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     2221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     2281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
     2341 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
     2401 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
     2461 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
     2521 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
     2581 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
     2641 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     2701 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     2761 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     2821 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     2881 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     2941 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     3001 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     3061 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     3121 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     3181 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     3241 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     3301 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     3361 taggcgccgg aattagatct ctcgaggcca ccatggacta caaagacgat gacgacaaga
     3421 tggctctacc ccggccaagt gaggccgtgc ctcaggacaa ggtgtgctac ccgccggaga
     3481 gcagcccgca gaacctggcc gcgtactaca cgcctttccc gtcctatgga cactacagaa
     3541 acagcctggc caccgtggag gaagacttcc aacctttccg gcagctggag gccgcagcgt
     3601 ctgctgcccc cgccatgccc cccttcccct tccggatggc gcctcccttg ctgagcccgg
     3661 gtctgggcct acagagggag cctctctacg atctgccctg gtacagcaag ctgccaccgt
     3721 ggtacccaat tccccacgtc cccagggaag tgccgccctt cctgagcagc agccacgagt
     3781 acgcgggtgc cagcagtgaa gatctgggcc accaaatcat tggtggcgac aacgagagtg
     3841 gcccgtgttg tggacctgac actttaattc caccgccccc tgcggatgct tctctgttac
     3901 ctgaggggct gaggacctcc cagttattac cttgctcacc cagcaagcag tcagaggatg
     3961 gtcccaaacc ctccaaccaa gaagggaagt cccctgctcg gttccagttc acggaggagg
     4021 acctgcactt cgttctgtac ggggtcactc ccagcctgga gcacccagcc agcctgcacc
     4081 atgcgatttc aggcctcctg gtccccccag acagctctgg atctgattct cttcctcaaa
     4141 ctctggatga agactccctt caacttccag aaggtctatg cctcatgcag acggtgtttg
     4201 gtgaagtccc acattttggt gtgttctgca gtagttttat cgccaaagga gtcaggtttg
     4261 ggccctttca aggtaaagtg gtcaatgcca gtgaagtgaa gacctacgga gacaattctg
     4321 tgatgtggga gatctttgaa gatggtcatt tgagccactt tatagatgga aaaggaggta
     4381 cggggaactg gatgtcctat gtcaactgtg cccgcttccc caaggagcag aacctagttg
     4441 ctgtgcagtg tcaagggcat atattttatg agagctgcaa agagattcat cagaaccaag
     4501 agctccttgt gtggtatgga gactgctatg agaaatttct ggatattcct gtgagccttc
     4561 aggtcacaga gccggggaag cagccatctg ggccctctga agagtctgca gaaggctagg
     4621 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     4681 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     4741 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     4801 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     4861 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     4921 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     4981 tgggaagggg tgggtccggg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     5041 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     5101 catctccggg cctttcgacc tgcagcccaa gcttaccatg accgagtaca agcccacggt
     5161 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     5221 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     5281 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     5341 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     5401 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     5461 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     5521 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     5581 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     5641 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     5701 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg
     5761 cccgaccgaa aggagcgcac gaccccatgc atcgataaaa taaaagattt tatttagtct
     5821 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac
     5881 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta
     5941 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     6001 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag
     6061 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     6121 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     6181 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg
     6241 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6301 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag
     6361 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac
     6421 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga
     6481 cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac
     6541 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6601 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6661 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6721 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6781 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6841 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6901 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6961 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7021 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7081 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7141 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7201 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7261 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7321 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7381 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7441 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa
//
