LOCUS       sequence-424572-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Histone H3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6264)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pKG333
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..475
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    540..739
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             940..1356
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1286..1308
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1324..1340
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1391..1400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1397..1399,1400..1402,1403..2113)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1397..1399,1400..1402,1403..2113)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1442..1463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1703..1722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2050..2071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2159..2179
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2192..2236
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2847..2867)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2852..3181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3032..3167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3189..3587
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3756..4225
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4355..4372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4526..5114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4606..4625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5274..6065,6066..6134))
                     /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     5897..5916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6135..6239)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg
       61 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
      121 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt
      181 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
      241 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
      301 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc
      361 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
      421 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg
      481 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca
      541 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg
      601 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact
      661 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt
      721 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg
      781 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
      841 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt
      901 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc
      961 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta
     1021 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc
     1081 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc
     1141 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc
     1201 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct
     1261 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc
     1321 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccagt gtggtggtac
     1381 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1441 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1501 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1561 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1621 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1681 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1741 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1801 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1861 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1921 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1981 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2041 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2101 cgagctgtac aagtccggag gggatcggtg gagctccacc ggtggcggcc gctctagaga
     2161 gaatctttat tttcagggcg ccgccaaatt caaagaaacc gctgctgcta aattcgaacg
     2221 ccagcacatg gacagcggag gtggaggtac tagtctcgag atggctcgta cgaagcaaac
     2281 agctcggaaa tccaccggcg gtaaagcgcc acgcaagcag ctggctacca aggctgctcg
     2341 caagagcgcg ccggctaccg gcggcgtgaa aaagcctcac cgttaccgcc cgggcactgt
     2401 ggctctgcgc gagatccgcc gctaccaaaa gtcgaccgag ttgctgattc ggaagctgcc
     2461 gttccagcgc ctggtgcgag aaatcgccca agacttcaag accgatcttc gcttccagag
     2521 ctctgcggtg atggcgctgc aggaggcttg tgaggcctac ttggtagggc tctttgagga
     2581 cacaaacctt tgcgccatcc atgctaagcg agtgactatt atgcccaaag acatccagct
     2641 cgctcgccgc atccgtgggg aaagggcata agaattctgc agtcgacggt accgcgggcc
     2701 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     2761 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2821 ttcgccagca cagtggtcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2881 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2941 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3001 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3061 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3181 aaagcttcat ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg
     3241 ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta
     3301 gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag
     3361 aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg
     3421 cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc
     3481 tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag
     3541 ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagat cgataaaata
     3601 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     3661 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     3721 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     3781 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     3841 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3901 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3961 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     4021 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     4081 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     4141 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     4201 actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt tggttttttt
     4261 tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa cgcgcgggga
     4321 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5221 ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6241 acatttcccc gaaaagtgcc acct
//
