LOCUS       sequence-177199-        7623 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 7623)
  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 7623)
  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 sh2 DNMT1
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..233
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     407..426
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        501..605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(606..674,675..1466)
                     /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(824..843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1637..2225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2126..2145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             2296..2476
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2994..3582
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3100..3111
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3509..3529
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     ncRNA           complement(3660..3786)
                     /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(3858..3979)
                     /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(4029..4628)
                     /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(4112..4132)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4609..4628
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4643..5050)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    5187..5304
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5602..5646)
                     /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    5831..6064
                     /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    6561..6686
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             6733..6913
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(6931..7129)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6956..6976)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7131..7510
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(join(7522..7623,1..228))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7536..7623,1..48)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7590..7609)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
       61 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      121 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
      181 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagcgc
      241 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
      301 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
      361 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
      421 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
      481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1141 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2281 acatgaaacg ggccctgcta gagattttcc acactgacta aaagggtctg agggatctct
     2341 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
     2401 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
     2461 ctaaccagag agacccagta cagtccggat gcagctctcg ggccatgtga tgaaatgcta
     2521 ggcggctgtc aaacctccac tctaatactt ctctctccgg gtcatccatc ccatgcaggc
     2581 tcacagggtg taacaagcgg gtgttctctc cttcattggc ttcttctacc ttctcttgct
     2641 caactggtac tagcttgtag caccatccaa aggtcagtgg atatctgatc cctggccctg
     2701 gtgtgtagtt ctgccaatca gggaagtagc cttgtgtgtg gtagatccac agatcaagga
     2761 tatcttgtct tcgttgggag tgaattagcc cttccagtcc ccccttttct tttaaaaagt
     2821 ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa aggtacctga ggtgtgactg
     2881 gaaaacccac ctcctcctcc tcttgtgctt ctagccaggc acaatcagca ttggtagctg
     2941 ctgtattgct acttgtgatt gctccatgtt tttctaggtc tcgacgcgat taagcgggga
     3001 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg
     3061 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta
     3121 gcagaaggac gtcccgcgca gaatccaggt ggcaacacag gcgagcagcc atggaaagga
     3181 cgtcagcttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     3241 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa
     3301 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc
     3361 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga
     3421 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     3481 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     3541 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttagcgatcg cggccccagg
     3601 ggccaccggt atcgatacgc gtcgccgcgt gtttaaacgc attagtcttc caattgaaaa
     3661 aagtgattta atttatacca ttttaattca gctttgtaaa aatgtatcaa agagatagca
     3721 aggtattcag ttttagtaaa caagataatt gctcctaaag tagccccttg aattccgagg
     3781 cagtaggcat acaaacgctc tcatcgacaa gtcaggagtc ttcgacgtca cacttgtcga
     3841 tgagagcgtt tgtgcgctca ctgtcaacag caatatacct tctcgagcct tctgttgggt
     3901 taacctgaag aagtaatccc agcaagtgtt tccaagatgt gcaggcaacg attctgtaaa
     3961 gtactgaagc ctcattcaac atagtatatg tgctgccgaa gcgagcactt aacaaggctt
     4021 gcggccgctc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc
     4081 tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg
     4141 gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg
     4201 acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac
     4261 cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc
     4321 agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct
     4381 tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc
     4441 ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg
     4501 atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg
     4561 agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac
     4621 tcggtcatgg tggctctaga cccccgggcg actcagtctg tcggaggact ggcgcgccga
     4681 gtgaggggtt gtgagctctt ttatagagct cgggaagcag aagcgcgcga acagaagcga
     4741 gaagcaggct gattggttaa ttcaaataag gcgcagggtc atttcaggtc cttgggggag
     4801 cctggaaaca tctgatgggt cttaagaaac tgctgagggt tgggccatat ctggggacca
     4861 tctgttcttg gcctcgggcc ggggccgaaa ctgcggtgac catctgttct tggccccggg
     4921 ccggggccga aactgctcac cgcagatatc ctgtttggcc caacgttagc tattttcatg
     4981 tacccgccct tgatctgaac ttctctattc ttggtttggt atttttccat gccttgcaaa
     5041 atggcgttac tgcagctagc ttgccaaacc tacaggtggg gtctttcatt cccccctttt
     5101 tctggagact aaataaaatc ttttatttta tcggggctgc agttaattaa ccaaactgga
     5161 tctctgctgt ccctgtaata aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat
     5221 tctttagttt gtatgtctgt tgctattatg tctactattc tttcccctgc actgtacccc
     5281 ccaatccccc cttttctttt aaaattgtgg atgaatactg ccatttgtct gcagaattgg
     5341 cgcacgcagt gccgatccgt tcactaatcg aatggatctg tctctgtctc tctctccacc
     5401 ttcttcttct attccttcgg gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa
     5461 cgataatggt gaatatccct gcctaactct attcactata gaaagtacag caaaaactat
     5521 tcttaaacct accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt
     5581 tgttatgtta aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc
     5641 attcttttct tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt
     5701 gttaatttct ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt
     5761 attactccaa ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc
     5821 ccaaatcccc aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg
     5881 gagctgcttg atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct
     5941 cagcaaattg ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag
     6001 cgtcattgac gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc
     6061 tcctattccc actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc
     6121 tactcctaat ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc
     6181 cctcatatct cctcctccag gtctgaagat cagcggccgg ccgcttgctg tgcggtggtc
     6241 ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct
     6301 atcctttgat gcacacaata gagggttgct actgtattat ataatgatct aagttcttct
     6361 gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt
     6421 tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt
     6481 tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta
     6541 attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta
     6601 gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca
     6661 gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg
     6721 ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt
     6781 taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt
     6841 gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta
     6901 accagagaga cccagtacag gcaaaacgcg ctgcttatat agacctccca ccgtacacgc
     6961 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     7021 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     7081 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     7141 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     7201 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     7261 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     7321 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     7381 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     7441 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     7501 aatcaatgtc aacgcgagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     7561 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
     7621 tca
//
