LOCUS       sequence-469964-        8670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing WT MPG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8670)
  AUTHORS   Tang S, Stokasimov E, Cui Y, Pellman D
  TITLE     Breakage of cytoplasmic chromosomes by pathological DNA base 
            excision repair.
  JOURNAL   Nature. 2022 Jun;606(7916):930-936. doi: 10.1038/s41586-022-04767-1.
            Epub 2022 Apr 27.
  PUBMED    35477155
REFERENCE   2  (bases 1 to 8670)
  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: 
            "10.1038/s41586-022-04767-1"; journalName: "Nature"; date: 
            "2022-06"; volume: "606"; issue: "7916"; pages: "930-936"
COMMENT     Sequence Label: pHAGE Flag HA MPG
FEATURES             Location/Qualifiers
     source          1..8670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1000..1021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1036..1041,1042..1059,1060..1066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1074..1090
                     /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     1079..1101
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1098..1114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1098..1114
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1164..1183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1375..2008
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2055..2180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2677..2910
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3095..3139
                     /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    3436..3553
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3605..3908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3909..4112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4063..4083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4109..4133
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4209..4218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4218..4241
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4254..4280
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4296..4320
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5200..5224)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5237..5804
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5254..5806
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5398..5415)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5625..5644
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5818..6417
                     /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(5818..5837)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6314..6334
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6434..7022
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6487..6507)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6905..6916)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7097..7330
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7449..7467)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7535..7639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7640..7708,7709..8500)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7858..7877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga
      841 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      901 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag
      961 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1021 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1081 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc
     1141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1201 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
     1261 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
     1321 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag
     1381 ggctaattca ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag
     1441 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
     1501 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
     1561 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     1621 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
     1681 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
     1741 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
     1801 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     1861 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1921 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1981 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     2041 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     2101 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     2161 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     2221 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     2281 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     2341 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     2401 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     2461 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     2521 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     2581 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     2641 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     2701 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     2761 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     2821 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     2881 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     2941 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     3001 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     3061 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     3121 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     3181 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     3241 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     3301 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     3361 cagatccatt cgattagtga acggatctcg acggtatcgc cgaattaatt cacaaatggc
     3421 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     3481 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     3541 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttgga ctagtggagt
     3601 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3661 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3721 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3781 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3841 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3901 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3961 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4021 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4081 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     4141 gacgccatcc acgctgtttt gacctccata gaagacaccg gcggccgcag atctctcgag
     4201 gttgatctgc caccatggac tacaaggatg acgatgacaa gctcgatgga ggatacccat
     4261 acgatgttcc agattacgct gctcgaggtt atcaaacaag tttgtacaaa aaagttggca
     4321 tgcccgcgcg cagcggggcc cagttttgcc gacggatggg gcaaaagaag cagcgaccag
     4381 ctagagcagg gcagccacac agctcgtccg acgcagccca ggcacctgca gagcagccac
     4441 acagctcgtc cgatgcagcc caggcacctt gccccaggga gcgctgcttg ggaccgccca
     4501 ccactccggg cccataccgc agcatctatt tctcaagccc aaagggccac cttacccgac
     4561 tggggttgga gttcttcgac cagccggcag tccccctggc ccgggcattt ctgggacagg
     4621 tcctagtccg gcgacttcct aatggcacag aactccgagg ccgcatcgtg gagaccgagg
     4681 catacctggg gccagaggat gaagccgccc actcaagggg tggccggcag accccccgca
     4741 accgaggcat gttcatgaag ccggggaccc tgtacgtgta catcatttac ggcatgtact
     4801 tctgcatgaa catctccagc cagggggacg gggcttgcgt cttgctgcga gcactggagc
     4861 ccctggaagg tctggagacc atgcgtcagc ttcgcagcac cctccggaaa ggcaccgcca
     4921 gccgtgtcct caaggaccgc gagctctgca gtggcccctc caagctgtgc caggccctgg
     4981 ccatcaacaa gagctttgac cagagggacc tggcacagga tgaagctgta tggctggagc
     5041 gtggtcccct ggagcccagt gagccggctg tagtggcagc agcccgggtg ggcgtcggcc
     5101 atgcagggga gtgggcccgg aaacccctcc gcttctatgt ccggggcagc ccctgggtca
     5161 gtgtggtcga cagagtggct gagcaggaca cacaggcctg cccaactttc ttgtacaaag
     5221 tggttcgata acgaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat
     5281 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     5341 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     5401 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     5461 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     5521 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     5581 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     5641 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     5701 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     5761 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     5821 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5881 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5941 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6001 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6061 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6121 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6181 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6241 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6301 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6361 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatgc
     6421 atcgatggat cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6481 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     6541 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6601 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     6661 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     6721 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     6781 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     6841 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     6901 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     6961 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     7021 gcctgagatc ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     7081 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     7141 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     7201 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     7261 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     7321 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     7381 tgaatatcag agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag
     7441 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     7501 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     7561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     7621 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     7681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     7741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     7801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     7861 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     7921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     7981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     8041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     8101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     8161 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     8221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     8281 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     8341 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     8401 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     8461 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     8521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     8581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     8641 agaccccgta gaaaagatca aaggatcttc
//
