LOCUS       sequence-436412-        6191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector with ubiquitin variant with reverted mutations 
            L69P and V70L (i53-DM mutant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6191)
  AUTHORS   Canny MD, Moatti N, Wan LCK, Fradet-Turcotte A, Krasner D, 
            Mateos-Gomez PA, Zimmermann M, Orthwein A, Juang YC, Zhang W, 
            Noordermeer SM, Seclen E, Wilson MD, Vorobyov A, Munro M, Ernst A, 
            Ng TF, Cho T, Cannon PM, Sidhu SS, Sicheri F, Durocher D
  TITLE     Inhibition of 53BP1 favors homology-dependent DNA repair and 
            increases CRISPR-Cas9 genome-editing efficiency.
  JOURNAL   Nat Biotechnol. 2018 Jan;36(1):95-102. doi: 10.1038/nbt.4021. Epub 
            2017 Nov 27.
  PUBMED    29176614
REFERENCE   2  (bases 1 to 6191)
  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/nbt.4021"; 
            journalName: "Nat Biotechnol"; date: "2018-01"; volume: "36"; issue:
            "1"; pages: "95-102"
COMMENT     Sequence Label: pMX-HA-UbvG08_MUT-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..6191
                     /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"
     LTR             890..1359
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(1530..2243,2244..2246,2247..2249))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1533..2243,2244..2246,2247..2249))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1575..1596)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1924..1943
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2183..2204
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(2250..2258,2259..2261,2262..2837)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2423..2442)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2652..2669
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(3103..3129)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3161..3535
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3207..3223)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3545..3961)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3561..3577)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3593..3615)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4020..4377
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4440..5033
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5063..5167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5168..5236,5237..6028)
                     /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(5386..5405)
                     /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 tgcaactatg gccatttaat
      841 gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt gaaagacccc
      901 cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga gaccacaagt
      961 cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact cagtcaatcg
     1021 gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg ggagcagaag
     1081 cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca cagggtcatt
     1141 tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc tgagggctgg
     1201 accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag
     1261 atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag ccggggcagg
     1321 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcctgacc
     1381 ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa aatggcgtta
     1441 cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
     1501 taaataaaat cttttatttt atcgtcgact tacttgtaca gctcgtccat gccgagagtg
     1561 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     1621 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     1681 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     1741 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     1801 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1861 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1921 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     1981 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     2041 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     2101 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     2161 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     2221 gtgaacagct cctcgccctt gctcaccatg gttgtggcca tattatcatc gtgtttttca
     2281 aaggaaaacc acgtccccgt ggttcggggg gcctagacgt ttttttaacc tcgactaaac
     2341 acatgtaaag catgtgcacc gaggccccag atcagatccc atacaatggg gtaccttctg
     2401 ggcatccttc agccccttgt tgaatacgct tgaggagagc catttgactc tttccacaac
     2461 tatccaactc acaacgtggc actggggttg tgccgccttt gcaggtgtat cttatacacg
     2521 tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt
     2581 cgctacagac gttgtttgtc ttcaagaagc ttccagagga actgcttcct tcacgacatt
     2641 caacagacct tgcattcctt tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa
     2701 gacagggcca ggtttccggg ccctcacatt gccaaaagac ggcaatatgg tggaaaataa
     2761 catatagaca aacgcacacc ggccttattc caagcggctt cggccagtaa cgttaggggg
     2821 ggggggaggg agaggggggg gggggggcgg aatttacgta gcggccgctc aacgaagtct
     2881 caacactaga tgaagtttag agtcctttag aatattgtag tcagacaaag tacgtccatc
     2941 ttccagcgat ttgccagcaa aggccagtct ctgctgatca ggaggaattc cttccttatc
     3001 ctggatcttg gcctttacat tttctatcgt atccgagggt tcaacctcga gggtgatggt
     3061 tttcccggta agggttttca cgaaaatcaa cattgcagct gcagcgtaat ctggaacatc
     3121 gtatgggtac atggtggctt aattaactag ctagatccgg cagtctagag gatggtccac
     3181 ccccggggtc ggcagccttc acgtgggcgg cgtgtatcca agctgcgatg ccgtctactt
     3241 tgagggcggt gggggtggtc agcaggactg tgtaaggtcc tttccagcga ggttctaggt
     3301 tcttagtctg gtgtcggcgg acccacactg tgtcgccgac tcggtaaggg tgaggtacca
     3361 ccggtcggtc cagttgttct tggtaggctg ccgccagagg tctccagact tcgtgctgga
     3421 ctaagtagag agcctgtaag tgagcttgga gagaggggct gttagtaact cttgtcatgt
     3481 cagggtcagg gaagtttaca aggggcgggg gtgccccata taagatctca tatggccata
     3541 tgggggcgcc tagagaagga gtgagggctg gataaaggga ggatcgaggc ggggtcgaac
     3601 gaggaggttc aagggggaga gacggggcgg atggaggaag aggaggcgga ggcttagggt
     3661 gtacaaaggg cttgacccag ggaggggggt caaaagccaa ggcttcccag gtcacgatgt
     3721 aggggacctg gtctgggtgt ccatgcgggc caggtgaaaa gaccttgatc ttaacctggg
     3781 tgatgaggtc tcggttaaag gtgccgtctc gcggccatcc gacgttaaag gttggccatt
     3841 ctgcagagca gaaggtaacc caacgtctct tcttgacatc taccgactgg ttgtgagcga
     3901 tccgctcgac atctttccag tgacctaagg tcaaacttaa gggagtggta acagtctggc
     3961 cctaattttc agacaaatac agaaacacag tcagacagag acaacacaga acgatgctgc
     4021 agcagacaag acgcgcggcg cggcttcggt cccaaaccga aagcaaaaat tcagacggag
     4081 gcgggaactg ttttaggttc tcgtctccta ccagaaccac atatccctcc tattaggggg
     4141 gtgcaccaaa gagtccaaaa cgatcgggat ttttggactc aggtcgggcc acaaaaacgg
     4201 cccccgaagt ccctgggacg tctcccaggg ttgcggccgg gtgttccgaa ctcgtcagtt
     4261 ccaccacggg tccgccagat acagagctag ttagctaact agtaccgacg caggcgcata
     4321 aaatcagtca tagacactag acaatcggac agacacagat aagttgctgg ccagcttacc
     4381 tcccggtggt gggtcggtgg tccctgggca ggggtctccc gatcccggac gagcccccaa
     4441 atgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc aaggaacagc
     4501 gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt acccgggcga
     4561 ctcagtcaat cggaggactg gcgcgccgag tgaggggttg tgggctcttt tattgagctc
     4621 ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt tcaaataagg
     4681 cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc tctagaaact
     4741 gctgagggct ggaccgcatc tggggaccat ctgttcttgg ccctgagccg gggcaggaac
     4801 tgcttaccac agatatcctg tttggcccat attcagctgt tccatctgtt cttggccctg
     4861 agccggggca ggaactgctt accacagata tcctgtttgg cccatattca gctgttccat
     4921 ctgttcctga ccttgatctg aacttttcta ttctcagtta tgtatttttc catgccttgc
     4981 aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc attatgcagg
     5041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5701 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6061 gattgatttg cggccggccg caaaaggatc taggtgaaga tcctttttga taatctcatg
     6121 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6181 aaaggatctt c
//
