LOCUS       sequence-436413-        6190 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Retroviral vector with ubiquitin variant that binds to and occludes 
            the ligand binding site of the 53BP1 Tudor domain (i53).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6190)
  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 6190)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..6190
                     /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..2836)
                     /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(3102..3128)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3160..3534
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     complement(3206..3222)
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(3544..3960)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(3560..3576)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(3592..3614)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    4019..4376
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             4439..5032
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        5062..5166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5167..5235,5236..6027)
                     /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(5385..5404)
                     /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 gggggaggga gagggggggg ggggggcgga atttacgtag cggccgctca acgaagtctc
     2881 aacagaggat gaagtttaga gtcctttaga atattgtagt cagacaaagt acgtccatct
     2941 tccagcgatt tgccagcaaa ggccagtctc tgctgatcag gaggaattcc ttccttatcc
     3001 tggatcttgg cctttacatt ttctatcgta tccgagggtt caacctcgag ggtgatggtt
     3061 ttcccggtaa gggttttcac gaaaatcaac attgcagctg cagcgtaatc tggaacatcg
     3121 tatgggtaca tggtggctta attaactagc tagatccggc agtctagagg atggtccacc
     3181 cccggggtcg gcagccttca cgtgggcggc gtgtatccaa gctgcgatgc cgtctacttt
     3241 gagggcggtg ggggtggtca gcaggactgt gtaaggtcct ttccagcgag gttctaggtt
     3301 cttagtctgg tgtcggcgga cccacactgt gtcgccgact cggtaagggt gaggtaccac
     3361 cggtcggtcc agttgttctt ggtaggctgc cgccagaggt ctccagactt cgtgctggac
     3421 taagtagaga gcctgtaagt gagcttggag agaggggctg ttagtaactc ttgtcatgtc
     3481 agggtcaggg aagtttacaa ggggcggggg tgccccatat aagatctcat atggccatat
     3541 gggggcgcct agagaaggag tgagggctgg ataaagggag gatcgaggcg gggtcgaacg
     3601 aggaggttca agggggagag acggggcgga tggaggaaga ggaggcggag gcttagggtg
     3661 tacaaagggc ttgacccagg gaggggggtc aaaagccaag gcttcccagg tcacgatgta
     3721 ggggacctgg tctgggtgtc catgcgggcc aggtgaaaag accttgatct taacctgggt
     3781 gatgaggtct cggttaaagg tgccgtctcg cggccatccg acgttaaagg ttggccattc
     3841 tgcagagcag aaggtaaccc aacgtctctt cttgacatct accgactggt tgtgagcgat
     3901 ccgctcgaca tctttccagt gacctaaggt caaacttaag ggagtggtaa cagtctggcc
     3961 ctaattttca gacaaataca gaaacacagt cagacagaga caacacagaa cgatgctgca
     4021 gcagacaaga cgcgcggcgc ggcttcggtc ccaaaccgaa agcaaaaatt cagacggagg
     4081 cgggaactgt tttaggttct cgtctcctac cagaaccaca tatccctcct attagggggg
     4141 tgcaccaaag agtccaaaac gatcgggatt tttggactca ggtcgggcca caaaaacggc
     4201 ccccgaagtc cctgggacgt ctcccagggt tgcggccggg tgttccgaac tcgtcagttc
     4261 caccacgggt ccgccagata cagagctagt tagctaacta gtaccgacgc aggcgcataa
     4321 aatcagtcat agacactaga caatcggaca gacacagata agttgctggc cagcttacct
     4381 cccggtggtg ggtcggtggt ccctgggcag gggtctcccg atcccggacg agcccccaaa
     4441 tgaaagaccc ccgctgacgg gtagtcaatc actcagagga gaccctccca aggaacagcg
     4501 agaccacaag tcggatgcaa ctgcaagagg gtttattgga tacacgggta cccgggcgac
     4561 tcagtcaatc ggaggactgg cgcgccgagt gaggggttgt gggctctttt attgagctcg
     4621 gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt caaataaggc
     4681 acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct ctagaaactg
     4741 ctgagggctg gaccgcatct ggggaccatc tgttcttggc cctgagccgg ggcaggaact
     4801 gcttaccaca gatatcctgt ttggcccata ttcagctgtt ccatctgttc ttggccctga
     4861 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc
     4921 tgttcctgac cttgatctga acttttctat tctcagttat gtatttttcc atgccttgca
     4981 aaatggcgtt acttaagcta gcttgccaaa cctacaggtg gggtctttca ttatgcaggt
     5041 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5101 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5161 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5221 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5281 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5341 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5401 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5461 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5521 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5581 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5641 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5701 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     5761 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5821 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5881 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5941 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6001 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6061 attgatttgc ggccggccgc aaaaggatct aggtgaagat cctttttgat aatctcatga
     6121 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6181 aaggatcttc
//
