LOCUS       sequence-426798-        8241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-HMBG1 in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8241)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 8241)
  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: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: mCherry-HMBG1
FEATURES             Location/Qualifiers
     source          1..8241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1642..1945
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1946..2149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2188..2197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2194..2901
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2339..2357)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2607..2626
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3636..4135
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3671..3694)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4068..4087
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4156..4755
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4156..4175)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4652..4672
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4940..5372
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5475..5497)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5633..5653)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5638..5967
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5818..5953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5989..6008
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6089..6106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6260..6848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6340..6359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7019..7810,7811..7879))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7642..7661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7880..7984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8052..8070
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8170..8241,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc atagttatta atagtaatca attacggggt cattagttca
     1621 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1681 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1741 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1801 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1861 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1921 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1981 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2041 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2101 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2161 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggaggataac
     2221 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2281 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2341 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2401 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2461 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2521 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2581 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2641 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2701 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2761 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2821 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2881 ggcatggacg agctgtacaa gtccggactc agatctggag gcggaggaag cggcaaagga
     2941 gatcctaaga agccgagagg caaaatgtca tcatatgcat tttttgtgca aacttgtcgg
     3001 gaggagcata agaagaagca cccagatgct tcagtcaact tctcagagtt ttctaagaag
     3061 tgctcagaga ggtggaagac catgtctgct aaagagaaag gaaaatttga agatatggca
     3121 aaggcggaca aggcccgtta tgaaagagaa atgaaaacct atatccctcc caaaggggag
     3181 acaaaaaaga agttcaagga tcccaatgca cccaagaggc ctccttcggc cttcttcctc
     3241 ttctgctctg agtatcgccc aaaaatcaaa ggagaacatc ctggcctgtc cattggtgat
     3301 gttgcgaaga aactgggaga gatgtggaat aacactgctg cagatgacaa gcagccttat
     3361 gaaaagaagg ctgcgaagct gaaggaaaaa tacgaaaagg atattgctgc atatcgagct
     3421 aaaggaaagc ctgatgcagc aaaaaaggga gttgtcaagg ctgaaaaaag caagaaaaag
     3481 aaggaagagg aggaagatga ggaagatgaa gaggatgagg aggaggagga agatgaagaa
     3541 gatgaagatg aagaagaaga tgatgatgat gaataagtcg acggtaccgc gggcccggga
     3601 tccaccggat ctagataact gatcataatt ctaccgggta ggggaggcgc ttttcccaag
     3661 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     3721 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     3781 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     3841 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     3901 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     3961 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     4021 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     4081 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     4141 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     4201 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     4261 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     4321 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     4381 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     4441 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     4501 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     4561 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     4621 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     4681 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4741 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     4801 ccccatgcat cggcgtctcg atcgagatat cagtggtcca ggctctagtt ttgactcaac
     4861 aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag attttattta
     4921 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctactagctt
     4981 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat
     5041 caaggtcagg aacagatgga acagggtcga tcgaccctag agaaccatca gatgtttcca
     5101 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     5161 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5221 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     5281 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     5341 gattgactac ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc
     5401 ctgcccaggg accaccgacc caccaccggg aggtaagctg gctgcctcgc gcgtttcggt
     5461 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5521 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5581 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt agatccggct gtggaatgtg
     5641 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5701 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5761 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5821 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5881 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5941 ttttttggag gcctaggctt ttgcaaaaag cttactggct taactatgcg gcatcagagc
     6001 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6061 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6121 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6181 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6241 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6301 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6361 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6421 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6481 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6541 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6601 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6661 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6721 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6781 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6841 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6901 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6961 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7021 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7081 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7141 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7201 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7261 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7321 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7381 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7441 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7501 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7561 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7621 cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7681 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7741 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7801 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7861 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7921 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7981 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     8041 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaatta gcttggccat
     8101 tgcatacgtt gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
     8161 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     8221 ttcatagccc atatatggag t
//
