LOCUS       sequence-258848-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of flag-tagged mouse TRIB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  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 7581)
  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-TRIB1
FEATURES             Location/Qualifiers
     source          1..7581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    50..391
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             458..874
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     804..826
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     842..858
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             890..913
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    2043..2610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2060..2612
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2204..2221)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2431..2450
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2624..2626,2627..2629,2630..3343)
                     /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(2669..2690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2930..2949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3277..3298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3445..3959
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4121..4137
                     /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(4145..4151,4152..4169,4170..4175))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4190..4211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4328..4345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4499..5087)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4579..4598)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5258..6049,6050..6118))
                     /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     5881..5900
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6119..6223)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6291..6309
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6347..6369)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6469..6488
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6682..6704
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6832..6851
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             7051..7567
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 agatttggag acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag
       61 cggtcgtttc gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg
      121 cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga
      181 cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt
      241 ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt
      301 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
      361 ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct
      421 ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt
      481 aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga
      541 tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg
      601 atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt
      661 ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt
      721 ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct
      781 tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc
      841 cctttatcca gccctcactc cttctctagg cgccggaatt agatccatgg actacaaaga
      901 cgatgacgac aagatgcggg tcggtcccgt gcgttttgcc ctgagcggcg cctcgcagcc
      961 ccgcggtccg ggtctactgt ttccggctgc ccggggaacc ccggccaaac gcctgctgga
     1021 cacggacgac gcgggggcag tggcggccaa gtgtccgcgc ctctctgagt gctcgagtcc
     1081 ccccgactac ctcagtcccc ctggctcgcc ctgcagccct cagcctccgc cctccacgca
     1141 ggggaccggc ggcagctgtg tgagctcacc cgggcccagc cgaatcgccg actacctgct
     1201 gctaccccta gctgagcgcg agcatgtgtc ccgggcactg tgcatccaca ccggccgcga
     1261 gctgcgctgc aaggaattcc ccattaaaca ctaccaggac aaaatcaggc cttacatcca
     1321 gctgccgtcc catagcaaca tcactggcat tgtggaggtg ctccttggtg agagcaaggc
     1381 ctacgtcttc ttcgagaagg actttggaga catgcactcc tatgtgcgga gccgaaaacg
     1441 gcttcgggaa gaggaggccg ctcggctctt caagcagatt gtttccgccg tcgcccactg
     1501 ccaccagtcg gccatcgtgc tgggggacct gaagctcagg aagttcgtct tctccaccga
     1561 ggagagaacc cagcttagac tggaaagcct ggaagatact cacatcatta agggtgaaga
     1621 tgatgcgctg tcagataaac atggctgccc agcctacgtg agccccgaga tcctcaacac
     1681 tactgggacc tactctggaa aggcggcgga cgtctggagc ctcggggtga tgctctatac
     1741 gcttttggtc ggacgatacc cctttcatga ctcagatccg agtgcccttt tttccaaaat
     1801 ccgccgtgga cagttctgca ttcctgagca cgtctccccc aaagccaggt gcctcatccg
     1861 aagcctccta agacgagaac cttctgagag actcacagct cctcagatct tactccatcc
     1921 ctggttcgag tatgtcttgg aaccgggcta tgttgactca gaaataggaa cttcagacca
     1981 gattgttcct gagtaccagg aggacagtga cataagttcc ttcttctgct aggttaacga
     2041 attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     2101 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     2161 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     2221 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     2281 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     2341 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     2401 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     2461 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     2521 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     2581 gtggttttcc tttgaaaaac acgatgataa tatggccaca accatggtga gcaagggcga
     2641 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2701 caagttcagc gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2761 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2821 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2881 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2941 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3001 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3061 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt
     3121 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3181 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3241 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3301 cgccgccggg atcactctcg gcatggacga gctgtacaag taatgaatta attaagaatt
     3361 atcaagctta tcgataccgt cgacctgcag ccaagcttat cgataaaata aaagatttta
     3421 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3481 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     3541 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     3601 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     3661 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     3721 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     3781 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     3841 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     3901 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     3961 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     4021 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     4081 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     4141 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4201 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4261 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4681 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4741 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4801 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4861 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4921 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4981 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5041 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5101 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5521 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5581 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5641 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5701 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5761 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5821 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5881 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5941 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6001 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6061 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6121 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6181 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6241 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     6301 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     6361 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     6421 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     6481 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6541 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     6601 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     6661 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg
     6721 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     6781 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     6841 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     6901 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt
     6961 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa
     7021 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     7081 gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag
     7141 ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg
     7201 taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc
     7261 ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac
     7321 cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg
     7381 ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag
     7441 actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc atccgaatcg
     7501 tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac ctcgggggtc
     7561 tttcatttgg aggttccacc g
//
