LOCUS       sequence-217050-        6684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector designed to knock down MePCE and LARP7 
            simultaneously..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6684)
  AUTHORS   Shelton SB, Shah NM, Abell NS, Devanathan SK, Mercado M, Xhemalce B
  TITLE     Crosstalk between the RNA Methylation and Histone-Binding Activities
            of MePCE Regulates P-TEFb Activation on Chromatin.
  JOURNAL   Cell Rep. 2018 Feb 6;22(6):1374-1383. doi: 
            10.1016/j.celrep.2018.01.028.
  PUBMED    29425494
REFERENCE   2  (bases 1 to 6684)
  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.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-6- 6"; volume: "22"; issue: "6"; pages: "1374-1383"
COMMENT     Sequence Label: pSUPER.retro.puro-shMePCE_shLARP7
FEATURES             Location/Qualifiers
     source          1..6684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        33..247
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     168..187
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        334..582
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     334..354
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     505..524
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(663..1079)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(679..695)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(711..733)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    1146..1487
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1551..2067
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2267..2286)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2414..2436)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2630..2649)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2749..2771
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2809..2827)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2895..2999
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3000..3068,3069..3860)
                     /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(3218..3237)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4031..4619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4520..4539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4773..4790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4907..4928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4943..4948,4949..4966,4967..4973)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4981..4997
                     /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)."
     CDS             complement(5583..6182)
                     /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(5666..5686)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6163..6182
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(6207..6684,1..22))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6255..6274)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6648..6671
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gggaaaagcg cctcccctac ccggtagaat tcgaacgctg acgtcatcaa cccgctccaa
       61 ggaatcgcgg gcccagtgtc actaggcggg aacacccagc gcgcgtgcgc cctggcagga
      121 agatggctgt gagggacagg ggagtggcgc cctgcaatat ttgcatgtcg ctatgtgttc
      181 tgggaaatca ccataaacgt gaaatgtctt tggatttggg aatcttataa gttctgtatg
      241 agaccacaga tcaagccaga gcagttcagt tccttcaaga gaggaactga actgctctgg
      301 ctttttaagc ttatcgataa ggtcgggcag gaagagggcc tatttcccat gattccttca
      361 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
      421 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
      481 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
      541 gatttcttgg gtttatatat cttgtggaaa ggacgatcgt ccaagttaat caccaaagct
      601 gttcaagaga cagctttggt gattaacttt ttttgtcgac ctcgagagat cgatctaatt
      661 ccggcgccta gagaaggagt gagggctgga taaagggagg atcgaggcgg ggtcgaacga
      721 ggaggttcaa gggggagaga cggggcggat ggaggaagag gaggcggagg cttagggtgt
      781 acaaagggct tgacccaggg aggggggtca aaagccaagg cttcccaggt cacgatgtag
      841 gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt aacctgggtg
      901 atgaggtctc ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt tggccattct
      961 gcagagcaga aggtaaccca acgtctcttc ttgacatcta ccgactggtt gtgagcgatc
     1021 cgctcgacat ctttccagtg acctaaggtc aaacttaagg gagtggtaac agtctggccc
     1081 taattttcag acaaatacag aaacacagtc agacagagac aacacagaac gatgctgcag
     1141 cgctgcagca gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa ttcagacgga
     1201 ggcgggaact gttttaggtt ctcgtctcct accagaacca catatcctga cggggtcgga
     1261 ttccacatcg actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa gtccctggga
     1321 cgtctcccag ggttgcggcc gggtgttcag aactcgtcag ttccaccacg ggtccgccag
     1381 atacagagct agttagctaa ctagtacaga cgcaggcgca aacattagat gccggcacaa
     1441 acacgcacaa agacagagac agacacgaaa cgaccgctgg ccagcttacc tcccggcggg
     1501 ggggtcggtg gtccctgggc aggggtctcc aaatctcggt ggaacctcca aatgaaagac
     1561 ccccgaggtg ggcagtcaat caatctgagg agaccctccc aaggatcagc gagtccacga
     1621 ttcggatgca aacagcaaga ggctttattg ggaatacggg tacccgggcg acgcagtcta
     1681 tcggaggact ggcgcgccga gtgaggggtt gtgggctctt ttattgagct cggggagcag
     1741 aagcgcgcga acagaagcga gaagcgaact gattggttag ttcaaataag gcacagggtc
     1801 atttcaggtc cttggggcac cctggaaaca tctgatggtt ctctagaaac tgctgagggc
     1861 gggaccgcat ctggggacca tctgttcttg gccctgagcc ggggcaggaa ctgcttacca
     1921 cagatatcct gtttggccca tattctgctg tctctctgtt cctaaccttg atctgaactt
     1981 ctctattctc agttatgtat tttccatgcc ttgcaaaatg gcgttactta agctagcttg
     2041 ccaaacctac aggtggggtc tttcattccc ccctttttct ggagactaaa taaaatcttt
     2101 tattttatct atggctcgta ctctataggc ttcagctggt gatattgttg agtcaaaact
     2161 agagcctgga ccactgatat cctgtcttta acaaattgga ctaatcgcct ctacgccgga
     2221 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     2281 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     2341 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     2401 ttccttgcgc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2461 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2521 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     2581 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     2641 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2701 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2761 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2821 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2881 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2941 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3001 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3061 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3121 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3181 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3241 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3301 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3361 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3421 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3481 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3541 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3601 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3661 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3721 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3781 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3841 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3901 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3961 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4021 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4081 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4141 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4201 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4261 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4321 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4381 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4441 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4501 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4561 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4621 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4681 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4741 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4801 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     4861 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     4921 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     4981 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa
     5041 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac
     5101 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat
     5161 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
     5221 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg
     5281 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg
     5341 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
     5401 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagcttgcca
     5461 aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa aatcttttat
     5521 tttatcgatg catggggtcg tgcgctcctt tcggtcgggc gctgcgggtc gtggggcggg
     5581 cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     5641 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     5701 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
     5761 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     5821 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     5881 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     5941 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     6001 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     6061 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     6121 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     6181 atggtaagct agcttgggct gcaggtcgaa aggcccggag atgaggaaga ggagaacagc
     6241 gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga ccttcgggcg
     6301 cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct tcccagcctc
     6361 tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag gcctacccgc
     6421 ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc tacttccatt
     6481 tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac taggggagga
     6541 gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct accggtggat
     6601 gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag cggggctgct
     6661 aaagcgcatg ctccagactg cctt
//
