LOCUS       sequence-255657-        7521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of flag-tagged mouse TRIB3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7521)
  AUTHORS   Dedhia PH, Keeshan K, Uljon S, Xu L, Vega ME, Shestova O, 
            Zaks-Zilberman M, Romany C, Blacklow SC, Pear WS
  TITLE     Differential ability of Tribbles family members to promote 
            degradation of C/EBPalpha and induce acute myelogenous leukemia.
  JOURNAL   Blood. 2010 Aug 26;116(8):1321-8. doi: 10.1182/blood-2009-07-229450.
            Epub 2010 Apr 21.
  PUBMED    20410507
REFERENCE   2  (bases 1 to 7521)
  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-2009-07-229450"; journalName: "Blood"; date: 
            "2010-08-26- 26"; volume: "116"; issue: "8"; pages: "1321-8"
COMMENT     Sequence Label: MigR1-Flag-TRIB3
FEATURES             Location/Qualifiers
     source          1..7521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    170..511
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             578..994
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     924..946
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     962..978
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1010..1033
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2103..2670
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2120..2672
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2264..2281)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2491..2510
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2684..2686,2687..2689,2690..3403)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2729..2750)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2990..3009)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3337..3358
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3505..4019
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4181..4197
                     /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(4205..4211,4212..4229,4230..4235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4250..4271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4388..4405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4559..5147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4639..4658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5318..6109,6110..6178))
                     /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     5941..5960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6179..6283)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6351..6369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6407..6429)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6529..6548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6742..6764
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6892..6911
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7111..7521,1..106)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga
       61 gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg
      121 agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag
      181 cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg
      241 cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
      301 cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt
      361 ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt
      421 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
      481 ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct
      541 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
      601 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
      661 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
      721 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
      781 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
      841 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
      901 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
      961 cctttatcca gccctcactc cttctctagg cgccggaatt agatctatgg actacaaaga
     1021 cgatgacgac aagatgcgag ctacacctct ggctgcttct gctgatgttt cctgcaggaa
     1081 gaaaccgttg gagtttgatg acaatattga tgccaagtgt ccagtcctaa aacgagtgag
     1141 agatgagcct gagcccggac cactccccag cctgctgccc cccagcccac ctcccgcctc
     1201 agacttgtca cctgctgtgg cccctgcaac tcgactgggg ccttatatcc ttttggaacg
     1261 agagcaaggc agctgcagct atcgagccct gcactgcccc acaggcacag agtacacctg
     1321 caaggtgtac cctgccagcg aggcccaggc ggtgctggca ccttatgccc ggctgcctac
     1381 ccaccagcat gtggcccgtc ccacagaggt cctgctgggc tctcggctcc tttacatctt
     1441 tttcacgaag acccatgggg acttgcacag cctggtgcgc agccgccgcg gtatcccaga
     1501 gtccgaggct gccgggctct tccggcagat ggctagtgcc gtggcacact gccacaagca
     1561 cgggcttgtc ttgcgcgacc tcaagctgcg tcgctttgtc ttcagcacct gtgagaggac
     1621 gaagctggtg ctggagaacc tggaagatgc ctgcgtgatg actggatcag atgactctct
     1681 gtgggacaag catgcgtgcc ctgcctacgt gggaccagag atactcagct cccggccatc
     1741 ctactctggc aaagcggctg atgtctggag cctgggtgtg gcgctcttca ccatgctggc
     1801 tggcagatac ccattccacg actctgagcc agtcctgctc tttggcaaga tccgtagagg
     1861 gacctttgcc ctgcctgagg gcctatcagc cccagcccgc tgtctgatcc gctgtctcct
     1921 ccgcaaggaa ccttcagagc gacttgtggc cctgggaatc ctcttgcatc cctggttgag
     1981 agaggatcac ggccgagtct ctcctccaca gtctgaccga agggagatgg accaggtggt
     2041 cccagatggg ccacagctgg aggaggctga ggaaggggag gtggggctgt acggctagga
     2101 attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2161 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2221 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2281 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2341 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2401 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2461 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2521 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2581 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2641 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     2701 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2761 caagttcagc gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2821 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2881 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2941 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3001 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3061 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3121 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt
     3181 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3241 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3301 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3361 cgccgccggg atcactctcg gcatggacga gctgtacaag taatgaatta attaagaatt
     3421 atcaagctta tcgataccgt cgacctgcag ccaagcttat cgataaaata aaagatttta
     3481 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3541 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     3601 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     3661 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     3721 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3781 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3841 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3901 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     3961 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     4021 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     4081 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     4141 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     4201 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4261 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4321 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4381 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4441 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4501 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4561 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4981 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5041 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5161 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5221 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5281 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5341 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5401 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5461 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5521 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5581 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5641 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5701 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5761 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5821 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5881 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5941 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6001 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6061 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6121 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6181 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6241 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6301 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6361 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6421 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6481 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6541 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6601 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6661 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6721 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     6781 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6841 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6901 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6961 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     7021 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     7081 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     7141 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     7201 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     7261 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     7321 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     7381 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     7441 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     7501 actgcgtcgc ccgggtaccc g
//
