LOCUS       sequence-275091-        6145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid with H4 EGFP insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6145)
  AUTHORS   Swartz SZ, McKay LS, Su KC, Bury L, Padeganeh A, Maddox PS, Knouse 
            KA, Cheeseman IM
  TITLE     Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain 
            Centromere Identity and Proliferative Potential.
  JOURNAL   Dev Cell. 2019 Oct 7;51(1):35-48.e7. doi: 
            10.1016/j.devcel.2019.07.016. Epub 2019 Aug 15.
  PUBMED    31422918
REFERENCE   2  (bases 1 to 6145)
  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.devcel.2019.07.016"; journalName: "Dev Cell"; date: 
            "2019-10-7- 7"; volume: "51"; issue: "1"; pages: "35-48.e7"
COMMENT     Sequence Label: pLM033
FEATURES             Location/Qualifiers
     source          1..6145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             108..524
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     454..476
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     492..508
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             889..909
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             922..966
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             991..1881
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(1896..1916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1901..2230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2081..2216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2238..2636
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2805..3274
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3404..3421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3575..4163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3655..3674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4323..5114,5115..5183))
                     /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     4946..4965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5184..5288)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5319..5788
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5853..6052
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
       61 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
      121 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
      181 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
      241 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
      301 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
      361 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
      421 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
      481 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggccgg atccatgtct
      541 gggcgaggta aaggtggcaa ggggctgggt aagggaggcg ccaagcgcca ccggaaggtg
      601 ctgcgggaca atatccaagg cattacaaag ccggcgattc gccgtctcgc ccgacgtggg
      661 ggcgtcaagc gcatttctgg tctcatctac gaggagaccc ggggagtcct caaagtcttc
      721 ctggagaacg tgatccgtga cgcggtgact tacacggagc acgccaagcg caagaccgtc
      781 acggccatgg atgtggtgta cgcgctgaaa cgccagggtc gcacccttta tggtttcggc
      841 ggtgaattcg gggatcggtg gagctccacc ggtggcggcc gctctagaga gaatctttat
      901 tttcagggcg ccgccaaatt caaagaaacc gctgctgcta aattcgaacg ccagcacatg
      961 gacagcggag gtggaggtac tagtctcgac atggcagaaa tcggtactgg ctttccattc
     1021 gacccccatt atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc
     1081 gatggcaccc ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac
     1141 atcatcccgc atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc
     1201 aaatccgaca aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc
     1261 ttcatcgaag ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct
     1321 ctgggtttcc actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag
     1381 ttcatccgcc ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag
     1441 gccttccgca ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag
     1501 ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc
     1561 gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca
     1621 atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac
     1681 cagtcccctg tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc
     1741 gaagccgctc gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt
     1801 ctgaatctgc tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctt
     1861 tcgacgctcg agatttccgg ttgagtcgac cctgtggaat gtgtgtcagt tagggtgtgg
     1921 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1981 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2041 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2101 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2161 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2221 cttttgcaaa aagcttcatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     2281 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     2341 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     2401 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     2461 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     2521 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     2581 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagatc
     2641 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     2701 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     2761 gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     2821 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     2881 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     2941 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     3001 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     3061 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     3121 cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     3181 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     3241 gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca aaattaattt
     3301 ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga atcggccaac
     3361 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3421 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3481 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4081 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4141 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4261 cctagatcct tttgcggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     4321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4621 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4921 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5281 gttccgcgca catttccccg aaaagtgcca cctgcagcct gaatatgggc caaacaggat
     5341 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     5401 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     5461 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     5521 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     5581 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     5641 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     5701 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     5761 ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg ggagacccct
     5821 gcccagggac caccgaccca ccaccgggag gcaagctggc cagcaactta tctgtgtctg
     5881 tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta gttagctaac
     5941 tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac
     6001 cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag
     6061 tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa
     6121 cagttcccgc ctccgtctga atttt
//
