LOCUS       sequence-91469-e        7585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MSCV based expression vector - co-expresses GFP and murine IRF4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7585)
  AUTHORS   So AY, Sookram R, Chaudhuri AA, Minisandram A, Cheng D, Xie C, Lim 
            EL, Flores YG, Jiang S, Kim JT, Keown C, Ramakrishnan P, Baltimore D
  TITLE     Dual mechanisms by which miR-125b represses IRF4 to induce myeloid 
            and B-cell leukemias.
  JOURNAL   Blood. 2014 Aug 28;124(9):1502-12. doi: 
            10.1182/blood-2014-02-553842. Epub 2014 Jul 8.
  PUBMED    25006123
REFERENCE   2  (bases 1 to 7585)
  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: 
            "10.1182/blood-2014-02-553842"; journalName: "Blood"; date: 
            "2014-08-28- 28"; volume: "124"; issue: "9"; pages: "1502-12"
COMMENT     Sequence Label: pMIG-IRF4
FEATURES             Location/Qualifiers
     source          1..7585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    join(2790..3364,3365..3367,3368..3376)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2957..2974)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3184..3203
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3377..3379,3380..3382,3383..4096)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     RNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3422..3443)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3683..3702)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4030..4051
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4693..4709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4693..4709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4706..4728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4717..4733
                     /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(4741..4747,4748..4765,4766..4771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4786..4807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4924..4941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5095..5683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5175..5194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5854..6645,6646..6714))
                     /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     6477..6496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6715..6819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6887..6905
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6943..6965)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7065..7084
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7278..7300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7292..7309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7293..7309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7508..7527
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg aggccaccat gaacttggag
     1441 acgggcagcc ggggctcaga gttcggcatg agcgcagtga gctgcggcaa tgggaaactc
     1501 cgacagtggt tgatcgacca gatcgacagc ggcaagtacc ccgggctggt gtgggagaac
     1561 gaggagaaga gcgtcttccg catcccgtgg aaacacgcgg gcaagcagga ctacaatcgt
     1621 gaggaggacg ctgccctctt caaggcttgg gcattgttta aaggcaagtt ccgagaaggg
     1681 atcgacaagc cagatcctcc tacttggaag acaagattac gatgtgctct gaacaagagc
     1741 aatgactttg aggaattggt cgagaggagc cagctggata tctctgaccc atacaaggtg
     1801 tacaggattg ttccagaggg agccaaaaaa ggagcaaagc agctcacttt ggatgacaca
     1861 cagatggcca tgggccaccc ctaccccatg acagcacctt atggctctct gccagcccag
     1921 caggttcata actacatgat gccaccccat gacaggagct ggagggatta tgcccctgac
     1981 cagtcacacc cagaaatccc atatcaatgt cctgtgacgt ttggcccacg aggccaccac
     2041 tggcaaggcc catcttgtga aaatggttgc caggtgacag gaacctttta tgcttgtgcc
     2101 ccacctgagt cccaggctcc tggaatcccc attgagccaa gcataaggtc tgctgaagcc
     2161 ttggcgctct cagactgccg gctgcatatc tgcctgtatt accgggacat cctcgtgaaa
     2221 gagctgacca cgacgagccc tgaaggctgc cggatctccc acggacacac ctatgatgtt
     2281 agcaacctgg accaggtcct gtttccctac ccggacgaca atggacagag gaagaacatt
     2341 gagaagttgc tgagccacct ggagagggga ctggtcctct ggatggctcc agatgggctt
     2401 tatgccaaaa gactctgcca gagtaggatc tactgggatg ggcccctggc actgtgcagc
     2461 gatcggccca acaagctaga aagagaccag acttgcaagc tctttgacac acagcagttt
     2521 ctatcagagc tgcaagtgtt tgctcaccat ggccggccag caccgagatt ccaggtgact
     2581 ctgtgctttg gtgaggagtt tccagaccct cagagacaga ggaagctcat cacagctcat
     2641 gtggaacctc tgctagccag acaactgtat tactttgctc aacaaaacac tggacatttc
     2701 ctgaggggct acgagttacc tgaacacgtt accactccag attaccaccg ctccctccgt
     2761 cattcttcca tccaagagtg agaattccgc ccctctccct cccccccccc taacgttact
     2821 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     2881 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2941 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     3001 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     3061 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     3121 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     3181 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     3241 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     3301 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     3361 taatatggcc acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3421 cctggtcgag ctggacggcg acgtgaacgg ccacaagttc agcgtgtccg gcgagggcga
     3481 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3541 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3601 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3661 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3721 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3781 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3841 cgacaagcag aagaacggca tcaagcgcaa cttcaagatc cgccacaaca tcgaggacgg
     3901 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3961 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4021 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc acggcatgga
     4081 cgagctgtac aagtaagtcg acctgcagcc aagcttatcg ataaaataaa agattttatt
     4141 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     4201 agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa gttcagatca
     4261 aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
     4321 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
     4381 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaaatg accctgtgcc
     4441 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
     4501 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc
     4561 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
     4621 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcagta
     4681 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4741 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4801 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4861 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4921 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4981 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5041 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5101 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5161 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5221 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5281 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5341 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5401 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5461 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5521 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5581 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5641 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5701 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5761 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5821 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5881 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5941 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6001 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6061 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6121 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6181 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6241 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6301 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6361 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6421 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6481 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6541 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6601 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6661 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6721 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6781 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6841 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6901 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6961 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     7021 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     7081 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7141 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     7201 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     7261 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccacgctctc
     7321 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     7381 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     7441 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     7501 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     7561 cggccacgat gcgtccggcg tagag
//
