LOCUS       sequence-66430-e        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector encoding Tet-inducible mitochondrially targeted 
            Y147A mutant human Uracil-N-glycosylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  AUTHORS   Shokolenko IN, Wilson GL, Alexeyev MF
  TITLE     Persistent damage induces mitochondrial DNA degradation.
  JOURNAL   DNA Repair (Amst). 2013 Jul;12(7):488-99. doi: 
            10.1016/j.dnarep.2013.04.023. Epub 2013 May 27.
  PUBMED    23721969
REFERENCE   2  (bases 1 to 7418)
  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: 
            "10.1016/j.dnarep.2013.04.023"; journalName: "DNA Repair (Amst)"; 
            date: "2013-07"; volume: "12"; issue: "7"; pages: "488-99"
COMMENT     Sequence Label: pMA3287
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1035..1160
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1657..1890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2075..2119
                     /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    2413..2530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     complement(2629..2645)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2630..2646)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        2645..2959
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2653..2671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2689..2707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2724..2742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2760..2778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2796..2814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2831..2849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2867..2885
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2932..2956
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3073..3102
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     3794..3810
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3794..3810
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3859..3879)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3864..4193
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4044..4179
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4207..4806
                     /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(4207..4226)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4703..4723
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5424..5604
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5666..6254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5746..5765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6425..7216,7217..7285))
                     /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     7048..7067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7286..7390)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
      241 gcgttgacat tgattattga ctagctagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tagaccagat ctgagcctgg
      841 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      901 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      961 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     1021 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     1081 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     1141 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     1201 ttcggttaag gccaggggga aagaaaaaat ataaataaac atatagtatg ggcaagcagg
     1261 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     1321 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1381 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1441 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1501 ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1561 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1621 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1681 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1741 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1801 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1861 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1921 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1981 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     2041 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     2101 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     2161 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     2221 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     2281 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     2341 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattcgac aaatggcagt
     2401 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2461 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2521 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag taccgggccg
     2581 gcccgctcta gctagacgta ttaccgccag atcccccatc aagcttatcg ataccgtcga
     2641 cctcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
     2701 atagagaacg atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
     2761 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat
     2821 gtcgagttta tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     2881 agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
     2941 agtgaaccgt cagatcgcct ggagaattcc gccaccatgc tgtttaatct gaggattctg
     3001 ttaaacaatg cagcttttag aaatggtcac aacttcatgg ttcgaaattt tcggtgtgga
     3061 caaccactac aagagcagaa actcatcagc gaggaggacc tgggatctgc ccgcaacgtg
     3121 cccgtgggct ttggagagag ctggaagaag cacctcagcg gggagttcgg gaaaccgtat
     3181 tttatcaagc taatgggatt tgttgcagaa gaaagaaagc attacactgt ttatccaccc
     3241 ccacaccaag tcttcacctg gacccagatg tgtgacataa aagatgtgaa ggttgtcatc
     3301 ctgggacagg atccagccca tggacctaat caagctcacg ggctctgctt tagtgttcaa
     3361 aggcctgttc cgcctccgcc cagtttggag aacatttata aagagttgtc tacagacata
     3421 gaggattttg ttcatcctgg ccatggagat ttatctgggt gggccaagca aggtgttctc
     3481 cttctcaacg ctgtcctcac ggttcgtgcc catcaagcca actctcataa ggagcgaggc
     3541 tgggagcagt tcactgatgc agttgtgtcc tggctaaatc agaactcgaa tggccttgtt
     3601 ttcttgctct ggggctctta tgctcagaag aagggcagtg ccattgatag gaagcggcac
     3661 catgtactac agacggctca tccctcccct ttgtcagtgt atagagggtt ctttggatgt
     3721 agacactttt caaagaccaa tgagctgctg cagaagtctg gcaagaagcc cattgactgg
     3781 aaggagctgt gagtctagaa ctagtggatc ccccgggctg caggaattga tggccgccac
     3841 cgcggtggag gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3901 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3961 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4021 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4081 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4141 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctag
     4201 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4261 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4321 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4381 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4441 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4501 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4561 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4621 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4681 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4741 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4801 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
     4861 tcgatcaatt cgatggggga tcaattcgat taagtcgagg gtcgatcgac ctcgatcgag
     4921 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     4981 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     5041 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5101 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     5161 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
     5221 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
     5281 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
     5341 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
     5401 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5461 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5521 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5581 agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc
     5641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7381 gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
