LOCUS       sequence-358621-        8642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral eukaryotic expression vector with E.Coli Dam methylation 
            fused to mouse Emerin. Used in DamID experiments to methylate DNA 
            that interacts in the proximity of Emerin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8642)
  AUTHORS   Amendola M, van Steensel B
  TITLE     Nuclear lamins are not required for lamina-associated domain 
            organization in mouse embryonic stem cells.
  JOURNAL   EMBO Rep. 2015 May;16(5):610-7. doi: 10.15252/embr.201439789. Epub 
            2015 Mar 17.
  PUBMED    25784758
REFERENCE   2  (bases 1 to 8642)
  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: 
            "10.15252/embr.201439789"; journalName: "EMBO Rep"; date: "2015-05";
            volume: "16"; issue: "5"; pages: "610-7"
COMMENT     Sequence Label: pLenti GW V5 Eco/Dam mse EMD
FEATURES             Location/Qualifiers
     source          1..8642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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)."
     primer_bind     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     218..238
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        220..238
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        339..718
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        719..922
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     873..893
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             937..1117
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1164..1289
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1782..2015
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2200..2244
                     /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    2511..2628
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2667..2905
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     2852..2873
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     CDS             3800..3841
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    3857..3881
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4699..4723)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4751..5339
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4804..4824)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5222..5233)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5426..5659
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5768..5787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5822..5841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5892..6027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6047..6066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6048..6066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6076..6093)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6076..6092)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6085..6107)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6234..6689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6321..6340)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6531..6552
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6715..6819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6820..6888,6889..7680)
                     /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(7038..7057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7851..8439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8340..8359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8593..8610
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      241 acaaaagctg gagctgcaag cttggccatt gcatacgttg tatccatatc ataatatgta
      301 catttatatt ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta
      361 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      421 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      481 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      541 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      601 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      661 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      721 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      781 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      841 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      901 ggaggtctat ataagcagag ctcgtttagt gaaccggggt ctctctggtt agaccagatc
      961 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     1021 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     1081 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     1141 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     1201 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     1261 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     1321 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     1381 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     1441 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1501 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     1561 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     1621 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1681 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1741 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1801 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1861 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1921 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1981 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     2041 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     2101 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     2161 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     2221 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     2281 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     2341 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     2401 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     2461 agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga
     2521 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2581 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttat cgatcacgag
     2641 actagcctcg agagtaccct cgaccgccgg agtataaata gaggcgcttc gtctacggag
     2701 cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct aagcaaataa
     2761 acaagcgcag ctgaacaagc taaacaatct gcagtaaagt gcaagttaaa gtgaatcaat
     2821 taaaagtaac cagcaaccaa gtaaatcaac tgcaactact gaaatctgcc aagaagtaat
     2881 tattgaatac aagaagagaa ctctgaatac tttcaacaag ttaccgagaa agaagaactc
     2941 acacacagct agcgccacca tgaagaaaaa tcgcgctttt ttgaagtggg cagggggcaa
     3001 gtatcccctg cttgatgata ttaaacggca tttgcccaag ggcgaatgtc tggttgagcc
     3061 ttttgtaggt gccgggtcgg tgtttctcaa caccgacttt tctcgttata tccttgccga
     3121 tatcaatagc gacctgatca gtctctataa cattgtgaag atgcgtactg atgagtacgt
     3181 acaggccgca cgcgagctgt ttgttcccga aacaaattgc gccgaggttt actatcagtt
     3241 ccgcgaagag ttcaacaaaa gccaggatcc gttccgtcgg gcggtactgt ttttatattt
     3301 gaaccgctac ggttacaacg gcctgtgtcg ttacaatctg cgcggtgagt ttaacgtgcc
     3361 gttcggccgc tacaaaaaac cctatttccc ggaagcagag ttgtatcact tcgctgaaaa
     3421 agcgcagaat gcctttttct attgtgagtc ttacgccgat agcatggcgc gcgcagatga
     3481 tgcatccgtc gtctattgcg atccgcctta tgcaccgctg tctgcgaccg ccaactttac
     3541 ggcgtatcac acaaacagtt ttacgcttga acaacaagcg catctggcgg agatcgccga
     3601 aggtctggtt gagcgccata ttccagtgct gatctccaat cacgatacga tgttaacgcg
     3661 tgagtggtat cagcgcgcaa aattgcatgt cgtcaaagtt cgacgcagta taagcagcaa
     3721 cggcggcaca cgtaaaaagg tggacgaact gctggctttg tacaaaccag gagtcgtttc
     3781 acccgcgaaa aaaaccggtg gtaagcctat ccctaaccct ctcctcggtc tcgattctac
     3841 gatcgatttc gatcaaacaa gtttgtacaa aaaagcaggc tccgcggccg cccccttcac
     3901 catggacgac tatgcggttt tgtccgatac tgagctggcc gcagtgctac gccagtacaa
     3961 catccctcat gggcctattg tgggttccac tcgcaagctc tacgaaaaga aaatcttcga
     4021 gtacgagacc cagagaagga gactcttacc ccccaactcg tcatcttctt cattctccta
     4081 tcagttctca gacttggatt cagccgccgt ggactcagac atgtatgatc tgcccaaaaa
     4141 agaagacgcc ttactttacc agagcaagga ctataatgat gactactatg aggagagtta
     4201 tttgaccacc aagacatacg gggagcctga gtctgtgggc atgtccaaga gcttccgcca
     4261 gccggggacc tcacttgtag atgctgatac cttccatcac caggtgcgtg atgacatttt
     4321 ctcttcttta gaagaggaag gcaaggatag ggagcgcctc atctatggcc aagacagtgc
     4381 ctaccagagc atcgcacact accgccccat ttccaatgtc tccaggagct ctctgggctt
     4441 atcatattat cctacatcct ccacctcttc tgtgtcctcc tcttcatctt ctccctcttc
     4501 atggcttacc cgacgtgcca ttcggccaga gaagcaggcc cctgctgctg ctctgggcca
     4561 ggatcgacag gtcccactct ggggtcagct gttactcttc ctggtctttg ctgcctttct
     4621 gctctttgtt tactattcca tacaagctga agaaggcaac cccttctgga tggatcccta
     4681 aaaagggtgg gcgcgccgac ccagctttct tgtacaaagt ggttcgatcg aaccgcggtc
     4741 tagactcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4801 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4861 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     4921 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     4981 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5041 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5101 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     5161 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5221 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5281 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5341 tggaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5401 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5461 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5521 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5581 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5641 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5701 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6001 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6061 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6121 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6181 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6241 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6301 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6361 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6421 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6481 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6541 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6601 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6661 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     6721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7681 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7741 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7981 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8461 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8521 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8581 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8641 cc
//
