LOCUS       sequence-177198-        7622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing shRNA against murine DNMT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7622)
  AUTHORS   Leoni C, Montagner S, Rinaldi A, Bertoni F, Polletti S, Balestrieri 
            C, Monticelli S
  TITLE     Dnmt3a restrains mast cell inflammatory responses.
  JOURNAL   Proc Natl Acad Sci U S A. 2017 Feb 6. pii: 201616420. doi: 
            10.1073/pnas.1616420114.
  PUBMED    28167789
REFERENCE   2  (bases 1 to 7622)
  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. 2017 Feb 6. pii: 201616420. doi: 
            10.1073/pnas.1616420114."
COMMENT     Sequence Label: pAPM sh1 DNMT1
FEATURES             Location/Qualifiers
     source          1..7622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     186..206
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     380..399
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        474..578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(579..647,648..1439)
                     /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(797..816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1610..2198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2099..2118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             2269..2449
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2967..3555
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3073..3084
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3482..3502
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     ncRNA           complement(3633..3759)
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(3830..3951)
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     CDS             complement(4001..4600)
                     /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(4084..4104)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4581..4600
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4615..5022)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    5159..5276
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5574..5618)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5803..6036
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6533..6658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6705..6885
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6903..7101)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6928..6948)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7103..7482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(join(7494..7622,1..201))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7508..7622,1..21)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7562..7581)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
       61 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
      121 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      181 tccacaccct aactgacaca cattccacag cgcgtatatc tggcccgtac atcgcgaagc
      241 agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt
      301 gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc
      361 agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga
      421 tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa
      481 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
      541 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
      601 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
      661 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
      721 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
      781 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
      841 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
      901 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
      961 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1021 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1081 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1141 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1201 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1261 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1321 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1381 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     1441 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     1501 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     1561 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     1621 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1681 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1741 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1801 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1861 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1921 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1981 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2041 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2101 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2161 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2221 tacggttcct ggccttttgc tggccttttg ctcacatgaa acgggccctg ctagagattt
     2281 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     2341 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     2401 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacagtccg
     2461 gatgcagctc tcgggccatg tgatgaaatg ctaggcggct gtcaaacctc cactctaata
     2521 cttctctctc cgggtcatcc atcccatgca ggctcacagg gtgtaacaag cgggtgttct
     2581 ctccttcatt ggcttcttct accttctctt gctcaactgg tactagcttg tagcaccatc
     2641 caaaggtcag tggatatctg atccctggcc ctggtgtgta gttctgccaa tcagggaagt
     2701 agccttgtgt gtggtagatc cacagatcaa ggatatcttg tcttcgttgg gagtgaatta
     2761 gcccttccag tccccccttt tcttttaaaa agtggctaag atctacagct gccttgtaag
     2821 tcattggtct taaaggtacc tgaggtgtga ctggaaaacc cacctcctcc tcctcttgtg
     2881 cttctagcca ggcacaatca gcattggtag ctgctgtatt gctacttgtg attgctccat
     2941 gtttttctag gtctcgacgc gattaagcgg ggaggcggcc caaagggaga tccgactcgt
     3001 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     3061 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     3121 ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca acaccacgga
     3181 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     3241 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     3301 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     3361 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     3421 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     3481 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     3541 atccagaggt tgattagcga tcgcggcccc aggggccacc ggtatcgata cgcgtcgccg
     3601 cgtgtttaaa cgcattagtc ttccaattga aaaaagtgat ttaatttata ccattttaat
     3661 tcagctttgt aaaaatgtat caaagagata gcaaggtatt cagttttagt aaacaagata
     3721 attgctccta aagtagcccc ttgaattccg aggcagtagg catacaaacg ctctcatcga
     3781 caagttttct ccctcacaca ctcacttgtc gatgagagcg tttgtgcgct cactgtcaac
     3841 agcaatatac cttctcgagc cttctgttgg gttaacctga agaagtaatc ccagcaagtg
     3901 tttccaagat gtgcaggcaa cgattctgta aagtactgaa gcctcattca acatagtata
     3961 tgtgctgccg aagcgagcac ttaacaaggc ttgcggccgc tcaggcaccg ggcttgcggg
     4021 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     4081 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
     4141 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     4201 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     4261 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     4321 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     4381 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     4441 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg acggtgtggc
     4501 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg gggacgtcgt
     4561 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtggctcta gacccccggg
     4621 cgactcagtc tgtcggagga ctggcgcgcc gagtgagggg ttgtgagctc ttttatagag
     4681 ctcgggaagc agaagcgcgc gaacagaagc gagaagcagg ctgattggtt aattcaaata
     4741 aggcgcaggg tcatttcagg tccttggggg agcctggaaa catctgatgg gtcttaagaa
     4801 actgctgagg gttgggccat atctggggac catctgttct tggcctcggg ccggggccga
     4861 aactgcggtg accatctgtt cttggccccg ggccggggcc gaaactgctc accgcagata
     4921 tcctgtttgg cccaacgtta gctattttca tgtacccgcc cttgatctga acttctctat
     4981 tcttggtttg gtatttttcc atgccttgca aaatggcgtt actgcagcta gcttgccaaa
     5041 cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa tcttttattt
     5101 tatcggggct gcagttaatt aaccaaactg gatctctgct gtccctgtaa taaacccgaa
     5161 aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta
     5221 tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt
     5281 ggatgaatac tgccatttgt ctgcagaatt ggcgcacgca gtgccgatcc gttcactaat
     5341 cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc gggcctgtcg
     5401 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     5461 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     5521 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     5581 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     5641 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     5701 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     5761 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     5821 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     5881 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     5941 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     6001 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     6061 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     6121 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag
     6181 atcagcggcc ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct
     6241 tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg
     6301 ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct
     6361 gtcccagtat ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc
     6421 gttctagctc cctgcttgcc catactatat gttttaattt atattttttc tttccccctg
     6481 gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct
     6541 cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc
     6601 gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct
     6661 cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca
     6721 cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca
     6781 cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta
     6841 gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaacg
     6901 cgctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     6961 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     7021 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     7081 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     7141 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     7201 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     7261 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     7321 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     7381 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     7441 ccgtaattga ttactattaa taactagtca ataatcaatg tcaacgcgag ctttttgcaa
     7501 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
     7561 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     7621 ct
//
