LOCUS       sequence-217043-        6371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector designed to knock down MePCE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6371)
  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 6371)
  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
FEATURES             Location/Qualifiers
     source          1..6371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..228
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     149..168
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(296..312)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(297..313)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(331..747)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(347..363)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(379..401)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    814..1155
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             1219..1735
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(1935..1954)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2082..2104)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2298..2317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2417..2439
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2477..2495)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2563..2667
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2668..2736,2737..3528)
                     /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(2886..2905)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3699..4287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4188..4207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4441..4458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4575..4596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4611..4616,4617..4634,4635..4641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4649..4665
                     /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(5251..5850)
                     /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(5334..5354)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5831..5850
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(5875..6371,1..3))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5923..5942)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     6316..6339
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cccggtagaa ttcgaacgct gacgtcatca acccgctcca aggaatcgcg ggcccagtgt
       61 cactaggcgg gaacacccag cgcgcgtgcg ccctggcagg aagatggctg tgagggacag
      121 gggagtggcg ccctgcaata tttgcatgtc gctatgtgtt ctgggaaatc accataaacg
      181 tgaaatgtct ttggatttgg gaatcttata agttctgtat gagaccacag atcaagccag
      241 agcagttcag ttccttcaag agaggaactg aactgctctg gctttttaag cttatcgata
      301 ccgtcgacct cgagagatcg atctaattcc ggcgcctaga gaaggagtga gggctggata
      361 aagggaggat cgaggcgggg tcgaacgagg aggttcaagg gggagagacg gggcggatgg
      421 aggaagagga ggcggaggct tagggtgtac aaagggcttg acccagggag gggggtcaaa
      481 agccaaggct tcccaggtca cgatgtaggg gacctggtct gggtgtccat gcgggccagg
      541 tgaaaagacc ttgatcttaa cctgggtgat gaggtctcgg ttaaaggtgc cgtctcgcgg
      601 ccatccgacg ttaaaggttg gccattctgc agagcagaag gtaacccaac gtctcttctt
      661 gacatctacc gactggttgt gagcgatccg ctcgacatct ttccagtgac ctaaggtcaa
      721 acttaaggga gtggtaacag tctggcccta attttcagac aaatacagaa acacagtcag
      781 acagagacaa cacagaacga tgctgcagcg ctgcagcaga caagacgcgc ggcttcggtt
      841 ccaaaccgaa agcaaaaatt cagacggagg cgggaactgt tttaggttct cgtctcctac
      901 cagaaccaca tatcctgacg gggtcggatt ccacatcgac tcccttcctc aggtcgggcc
      961 acaaaaacgg cccccaaagt ccctgggacg tctcccaggg ttgcggccgg gtgttcagaa
     1021 ctcgtcagtt ccaccacggg tccgccagat acagagctag ttagctaact agtacagacg
     1081 caggcgcaaa cattagatgc cggcacaaac acgcacaaag acagagacag acacgaaacg
     1141 accgctggcc agcttacctc ccggcggggg ggtcggtggt ccctgggcag gggtctccaa
     1201 atctcggtgg aacctccaaa tgaaagaccc ccgaggtggg cagtcaatca atctgaggag
     1261 accctcccaa ggatcagcga gtccacgatt cggatgcaaa cagcaagagg ctttattggg
     1321 aatacgggta cccgggcgac gcagtctatc ggaggactgg cgcgccgagt gaggggttgt
     1381 gggctctttt attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga
     1441 ttggttagtt caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc
     1501 tgatggttct ctagaaactg ctgagggcgg gaccgcatct ggggaccatc tgttcttggc
     1561 cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata ttctgctgtc
     1621 tctctgttcc taaccttgat ctgaacttct ctattctcag ttatgtattt tccatgcctt
     1681 gcaaaatggc gttacttaag ctagcttgcc aaacctacag gtggggtctt tcattccccc
     1741 ctttttctgg agactaaata aaatctttta ttttatctat ggctcgtact ctataggctt
     1801 cagctggtga tattgttgag tcaaaactag agcctggacc actgatatcc tgtctttaac
     1861 aaattggact aatcgcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     1921 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     1981 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     2041 gttgggcgcc atctccttgc atgcaccatt ccttgcgccg tcgttttaca acgtcgtgac
     2101 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     2161 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     2221 ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2281 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2341 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2401 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2461 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     2521 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2581 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2641 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2701 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2761 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2821 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2881 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     2941 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3001 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3061 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3121 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3181 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3241 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3301 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3361 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3421 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3481 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3541 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3601 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3841 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4321 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4381 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4441 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4501 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4561 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     4621 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca caattctccc
     4681 cagctctttc tgcgctgtcc ataatgaact atttcaggag tattggagta tgtggattca
     4741 aaacagtggc taattgagtc aggattactt aatattcgac tcctaccctc agaatgttgt
     4801 tctccaactt caagaaactg ttacccatga aagacccccg ctgacgggta gtcaatcact
     4861 cagaggagac cctcccaagg aacagcgaga ccacaagtcg gatgcaactg caagagggtt
     4921 tattggatac acgggtaccc gggcgacgca gtctatcgga ggactggcgc gccgagtgag
     4981 gggttgtggg ctcttttatt gagctcgggg agcagaagcg cgcgaacaga agcgagaagc
     5041 gaactgattg gttagttcaa ataaggcaca gggtcatttc aggtccttgg ggcaccctgg
     5101 aaacatctga tggttctcta gcttgccaaa cctacaggtg gggtctttca ttcccccctt
     5161 tttctggaga ctaaataaaa tcttttattt tatcgatgca tggggtcgtg cgctcctttc
     5221 ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg tcatgcacca
     5281 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     5341 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
     5401 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     5461 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     5521 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     5581 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     5641 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
     5701 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
     5761 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
     5821 gaggcgcacc gtgggcttgt actcggtcat ggtaagctag cttgggctgc aggtcgaaag
     5881 gcccggagat gaggaagagg agaacagcgc ggcagacgtg cgcttttgaa gcgtgcagaa
     5941 tgccgggcct ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc ccctgagccc
     6001 gcccccggac ccaccccttc ccagcctctg agcccagaaa gcgaaggagc aaagctgcta
     6061 ttggccgctg ccccaaaggc ctacccgctt ccattgctca gcggtgctgt ccatctgcac
     6121 gagactagtg agacgtgcta cttccatttg tcacgtcctg cacgacgcga gctgcggggc
     6181 gggggggaac ttcctgacta ggggaggagt agaaggtggc gcgaaggggc caccaaagaa
     6241 cggagccggt tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt
     6301 gtagcgccaa gtgcccagcg gggctgctaa agcgcatgct ccagactgcc ttgggaaaag
     6361 cgcctcccct a
//
