LOCUS       sequence-173086-        8047 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral construct for expression of mouse Mdm2 cloned into the 
            EcoRI site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8047)
  TITLE     MSCV-mMdm2-IRES-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8047)
  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            "Unpublished"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: MSCV-mMDM2-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    545..886
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             953..1369
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1299..1321
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1337..1353
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    2973..3522
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3117..3134)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3344..3363
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3525..3527,3528..3530,3531..4244)
                     /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             join(3525..3527,3528..3530,3531..4241)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3570..3591)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3831..3850)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4178..4199
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4406..4920
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5082..5098
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5106..5112,5113..5130,5131..5136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5151..5172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5289..5306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5460..6048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5540..5559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6219..7010,7011..7079))
                     /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     6842..6861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7080..7184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7252..7270
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7308..7330)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7430..7449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7643..7665
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7793..7812
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8012..8047,1..481)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
       61 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
      121 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
      181 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
      241 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
      301 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
      361 tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggact
      421 cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg ggtctttcat
      481 ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc ccccgccggg
      541 aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg tttgtgccgg
      601 catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac
      661 ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga
      721 ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc
      781 gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
      841 atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat
      901 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
      961 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1021 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1081 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1141 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1201 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1261 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1321 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg gaattcgatt
     1381 cgatcaccgc gcttctcctg cggcctccag gccaatgtgc aataccaaca tgtctgtgtc
     1441 taccgagggt gctgcaagca cctcacagat tccagcttcg gaacaagaga ctctggttag
     1501 accaaaacca ttgcttttga agttgttaaa gtccgttgga gcgcaaaacg acacttacac
     1561 tatgaaagag attatatttt atattggcca gtatattatg actaagaggt tatatgacga
     1621 gaagcagcag cacattgtgt attgttcaaa tgatctccta ggagatgtgt ttggagtccc
     1681 gagtttctct gtgaaggagc acaggaaaat atatgcaatg atctacagaa atttagtggc
     1741 tgtaagtcag caagactctg gcacatcgct gagtgagagc agacgtcagc ctgaaggtgg
     1801 gagtgatctg aaggatcctt tgcaagcgcc accagaagag aaaccttcat cttctgattt
     1861 aatttctaga ctgtctacct catctagaag gagatccatt agtgagacag aagagaacac
     1921 agatgagcta cctggggagc ggcaccggaa gcgccgcagg tccctgtcct ttgatccgag
     1981 cctgggtctg tgtgagctga gggagatgtg cagcggcggc agcagcagca gtagcagcag
     2041 cagcagcgag tccacagaga cgccctcgca tcaggatctt gacgatggcg taagtgagca
     2101 ttctggtgat tgcctggatc aggattcagt ttctgatcag tttagcgtgg aatttgaagt
     2161 tgagtctctg gactcggaag attacagcct gagtgacgaa gggcacgagc tctcagatga
     2221 ggatgatgag gtctatcggg tcacagtcta tcagacagga gaaagcgata cagactcttt
     2281 tgaaggggat cctgagattt ccttagctga ctattggaag tgtacctcat gcaatgaaat
     2341 gaatcctccc cttccatcac actgcaaaag atgctggacc cttcgtgaga actggcttcc
     2401 agacgataag gggaaagata aagtggaaat ctctgaaaaa gccaaactgg aaaactcagc
     2461 tcaggcagaa gaaggcttgg atgtgcctga tggcaaaaag ctgacagaga atgatgctaa
     2521 agagccatgt gctgaggagg acagcgagga gaaggccgaa cagacgcccc tgtcccagga
     2581 gagtgacgac tattcccaac catcgacttc cagcagcatt gtttatagca gccaagaaag
     2641 cgtgaaagag ttgaaggagg aaacgcagga caaagacgag agtgtggaat ctagcttctc
     2701 cctgaatgcc atcgaaccat gtgtgatctg ccaggggcgg cctaaaaatg gctgcattgt
     2761 tcacggcaag actggacacc tcatgtcatg tttcacgtgt gcaaagaagc taaaaaaaag
     2821 aaacaagccc tgcccagtgt gcagacagcc aatccaaatg attgtgctaa cttacttcaa
     2881 ctagctgacc tgctcacaaa aatagaattt tatatcgata atcactagtg aattcgttaa
     2941 cctcgagcgg gatcaattcc gccccccccc taacgttact ggccgaagcc gcttggaata
     3001 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     3061 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     3121 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     3181 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     3241 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     3301 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     3361 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     3421 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     3481 gaaccacggg gacgtggttt tcctttgaaa aacacgataa taccatggtg agcaagggcg
     3541 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3601 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3661 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3721 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3781 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3841 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3901 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3961 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4021 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4081 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4141 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4201 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     4261 gagtcgacct gcaggcatgc aagcttcagg tagccggcta acgttaacaa ccggtacctc
     4321 tagaactata gctagcatgc gcaaatttaa agcgctgata tcgataaaat aaaagatttt
     4381 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     4441 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     4501 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     4561 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     4621 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4681 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4741 gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg
     4801 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4861 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
     4921 gggtaacagt ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt
     4981 aatcctgact caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt
     5041 cattatggac agcgcagaaa gagctgggga gaattgtgaa attgttatcc gctcacaatt
     5101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5881 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7201 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     7261 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     7321 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     7381 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     7441 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7501 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7561 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     7621 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc gcaaggaatg
     7681 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     7741 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     7801 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     7861 cggcgtagag gcgattagtc caatttgtta aagacaggat atcagtggtc caggctctag
     7921 ttttgactca acaatatcac cagctgaagc ctatagagta cgagccatag ataaaataaa
     7981 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     8041 agctagc
//
