LOCUS       sequence-271172-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid with H3.1A EGFP insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  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 6247)
  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: pLM032
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             292..312
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             325..369
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             394..1284
                     /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(1299..1319)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1304..1633
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1484..1619
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1641..2039
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             2208..2677
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2807..2824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2978..3566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3058..3077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3726..4517,4518..4586))
                     /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     4349..4368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4587..4691)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4722..5191
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5256..5455
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5656..6072
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6002..6024
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6040..6056
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 tatcagaagt ccactgaact gcttattcgt aaactacctt tccagcgcct ggtgcgcgag
       61 attgcgcagg actttaaaac agacctgcgt ttccagagct ccgctgtgat ggctctgcag
      121 gaggcgtgcg aggcctactt ggtagggcta tttgaggaca ctaacctgtg cgccatccac
      181 gccaagcgcg tcactatcat gcccaaggac atccagctcg cccgccgcat ccgcggagag
      241 agggcggaat tcggggatcg gtggagctcc accggtggcg gccgctctag agagaatctt
      301 tattttcagg gcgccgccaa attcaaagaa accgctgctg ctaaattcga acgccagcac
      361 atggacagcg gaggtggagg tactagtctc gacatggcag aaatcggtac tggctttcca
      421 ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg
      481 cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc
      541 aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg
      601 ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat
      661 gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc
      721 gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg
      781 gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc
      841 caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc
      901 gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac
      961 cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg
     1021 ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg
     1081 caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg
     1141 gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg
     1201 ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg
     1261 ctttcgacgc tcgagatttc cggttgagtc gaccctgtgg aatgtgtgtc agttagggtg
     1321 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1381 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1441 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1501 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1561 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1621 aggcttttgc aaaaagcttc atggccaagc ctttgtctca agaagaatcc accctcattg
     1681 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
     1741 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
     1801 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
     1861 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
     1921 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
     1981 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag
     2041 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     2101 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     2161 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     2221 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     2281 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2341 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     2401 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     2461 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     2521 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     2581 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     2641 tctgagtgat tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa
     2701 tttggttttt tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc
     2761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3661 tcacctagat ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct
     3721 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3781 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3841 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3901 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3961 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4021 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4081 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4141 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4201 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4261 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4321 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4381 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4441 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4501 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4561 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4621 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4681 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag
     4741 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga
     4801 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4861 agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc
     4921 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     4981 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     5041 cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa
     5101 ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag
     5161 tgattgacta cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc
     5221 cctgcccagg gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt
     5281 ctgtccgatt gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct
     5341 aactagctct gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc
     5401 aaccctggga gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg
     5461 gagtcgatgt ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta
     5521 aaacagttcc cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct
     5581 tgtctgctgc agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat
     5641 ttgtctgaaa attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg
     5701 aaagatgtcg agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt
     5761 accttctgct ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt
     5821 aaccgagacc tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac
     5881 ccagaccagg tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg
     5941 gtcaagccct ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc
     6001 ccccttgaac ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct
     6061 tctctaggcg ccggccggat ccaccatggc tcgcactaag caaactgctc ggaagtctac
     6121 tggtggcaag gcgccacgca aacagttggc cactaaggca gcccgcaaaa gcgctccggc
     6181 caccggcggc gtgaaaaagc cccaccgcta ccggccgggc accgtggctc tgcgcgagat
     6241 ccgccgt
//
