LOCUS       sequence-166323-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Negative control vector for pMKO.1 puro (Addgene plasmid #8452); 
            Contains shRNA against GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  AUTHORS   Masutomi K, Possemato R, Wong JM, Currier JL, Tothova Z, Manola JB, 
            Ganesan S, Lansdorp PM, Collins K, Hahn WC
  TITLE     The telomerase reverse transcriptase regulates chromatin state and 
            DNA damage responses.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 Jun 7. 102(23):8222-7.
  PUBMED    15928077
REFERENCE   2  (bases 1 to 6697)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2005 Jun 7. 102(23):8222-7."
COMMENT     Sequence Label: pMKO.1 puro GFP shRNA
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             205..380
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    443..642
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             843..1259
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1189..1211
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1227..1243
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1295..1543
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1295..1315
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1466..1485
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        1631..1960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1811..1946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1970..2569
                     /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(1970..1989)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2466..2486
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2765..3190
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(3293..3315)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3456..3785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3636..3771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3807..3826
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4198..4786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4278..4297)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4957..5748,5749..5817))
                     /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     5580..5599
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5818..5922)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6200..6218
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        6318..6697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcaataa aagagcccac aacccctcac tcggcgcgcc
      241 agtcttccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
      301 tgcatccgaa tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc
      361 cacgacgggg gtctttcatt tgggggctcg tccgggattt ggagacccct gcccagggac
      421 caccgaccca ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc
      481 tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta gttagctaac tagctctgta
      541 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
      601 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
      661 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
      721 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
      781 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
      841 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
      901 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
      961 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     1021 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     1081 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     1141 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     1201 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     1261 gaattgaaga tctgtcgaca aggtcgggca ggaagagggc ctatttccca tgattccttc
     1321 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
     1381 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
     1441 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
     1501 cgatttcttg gctttatata tcttgtggaa aggacgaaac accggctacg tccaggagcg
     1561 caccctcgag ggtgcgctcc tggacgtagc ctttttgaat tcgccagcac agtggtcgac
     1621 cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggcttcccag caggcagaag
     1681 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     1801 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1861 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     1921 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca tgaccgagta
     1981 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     2041 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
     2101 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     2161 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     2221 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     2281 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     2341 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     2401 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     2461 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     2521 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca
     2581 cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgatta gtccaatttg
     2641 ttaaagacag gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga
     2701 agcctataga gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg
     2761 ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca
     2821 aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg
     2881 gaacagggtc gaccctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat
     2941 gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt
     3001 ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
     3061 ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact
     3121 tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg
     3181 gtctttcatt tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca
     3241 ccaccgggag gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3301 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3361 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac
     3421 gtagcgatag cggagtgtag atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3721 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3781 gcaaaaagct tactggctta actatgcggc atcagagcag attgtactga gagtgcacca
     3841 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc
     3901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4021 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4081 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4861 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4921 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4981 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5041 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5101 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5161 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5221 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5281 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5341 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5401 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5461 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5521 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5581 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5641 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5701 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5761 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5821 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5881 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5941 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6001 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttaattatt gaagcattta
     6061 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6121 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6181 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattagc
     6241 ttggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     6301 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     6361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     6421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     6481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     6541 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     6601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     6661 gcagtacatc tacgtattag tcatcgctat taccatg
//
