LOCUS       sequence-151500-        6376 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNAi vector for Sirt6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  AUTHORS   Tennen RI, Bua DJ, Wright WE, Chua KF
  TITLE     SIRT6 is required for maintenance of telomere position effect in 
            human cells.
  JOURNAL   Nat Commun. 2011 Aug 16;2:433. doi: 10.1038/ncomms1443.
  PUBMED    21847107
REFERENCE   2  (bases 1 to 6376)
  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: "Nat 
            Commun."; date: "2011-08-16"; volume: "2:433. doi"; pages: " 
            10.1038/ncomms1443"
COMMENT     Sequence Label: pSUPERretro-Sirt6 shRNA1
FEATURES             Location/Qualifiers
     source          1..6376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     365..387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     515..534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             734..1250
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1314..1655
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1722..2138
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2068..2090
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2106..2122
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     2156..2172
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2157..2173
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2246..2460)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(2306..2325)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        2471..2970
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2506..2529)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2903..2922
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2995..3594
                     /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(2995..3014)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3491..3511
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    4180..4196
                     /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(4204..4210,4211..4228,4229..4234))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4249..4270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4387..4404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4558..5146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4638..4657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5317..6108,6109..6177))
                     /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     5940..5959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6178..6282)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6350..6368
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      241 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      301 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      361 ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc aaggagatgg
      421 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      481 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      541 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggcgattag
      601 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
      661 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
      721 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
      781 cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat caaggttagg
      841 aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
      901 ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca gtttctagag
      961 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     1021 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     1081 aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt cgcccgggta
     1141 cccgtattcc caataaagcc tcttgctgtt tgcatccgaa tcgtggactc gctgatcctt
     1201 gggagggtct cctcagattg attgactgcc cacctcgggg gtctttcatt tggaggttcc
     1261 accgagattt ggagacccct gcccagggac caccgacccc cccgccggga ggtaagctgg
     1321 ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc atctaatgtt
     1381 tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
     1441 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
     1501 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
     1561 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     1621 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
     1681 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
     1741 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1801 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1861 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1921 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1981 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     2041 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     2101 cctcccttta tccagccctc actccttctc taggcgccgg aattagatcg atctctcgag
     2161 gtcgacggta tcgataagct taaaaaaagc tggagcccaa ggaggaatct cttgaattcc
     2221 tccttgggct ccagcttggg gatctgtggt ctcatacaga acttataaga ttcccaaatc
     2281 caaagacatt tcacgtttat ggtgatttcc cagaacacat agcgacatgc aaatattgca
     2341 gggcgccact cccctgtccc tcacagccat cttcctgcca gggcgcacgc gcgctgggtg
     2401 ttcccgccta gtgacactgg gcccgcgatt ccttggagcg ggttgatgac gtcagcgttc
     2461 gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     2521 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     2581 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     2641 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     2701 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     2761 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     2821 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     2881 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     2941 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagctagct taccatgacc
     3001 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     3061 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     3121 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     3181 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     3241 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     3301 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     3361 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     3421 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     3481 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     3541 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     3601 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gataaaataa
     3661 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     3721 aagctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
     3781 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     3841 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     3901 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     3961 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg
     4021 gtaacagttt cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa
     4081 tcctgactca attagccact gttttgaatc cacatactcc aatactcctg aaatagttca
     4141 ttatggacag cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc
     4201 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4261 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4321 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4381 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4441 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4501 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4561 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4621 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4681 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4741 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4801 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4861 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4921 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4981 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5041 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5101 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5161 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5221 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5281 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5341 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5401 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5461 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5521 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5581 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5641 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5701 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5761 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5821 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5881 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5941 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6001 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6061 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6121 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6181 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6241 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6301 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6361 cacgaggccc tttcgt
//
