LOCUS       sequence-255677-        7509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of flag-tagged mouse TRIB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7509)
  AUTHORS   Keeshan K, He Y, Wouters BJ, Shestova O, Xu L, Sai H, Rodriguez CG, 
            Maillard I, Tobias JW, Valk P, Carroll M, Aster JC, Delwel R, Pear 
            WS
  TITLE     Tribbles homolog 2 inactivates C/EBPalpha and causes acute 
            myelogenous leukemia.
  JOURNAL   Cancer Cell. 2006 Nov;10(5):401-11. doi: 10.1016/j.ccr.2006.09.012.
  PUBMED    17097562
REFERENCE   2  (bases 1 to 7509)
  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: 
            "10.1016/j.ccr.2006.09"; journalName: "Cancer Cell"; date: 
            "2006-11"; volume: "10"; issue: "5"; pages: "401-11"
COMMENT     Sequence Label: MigR1-Flag-TRIB2
FEATURES             Location/Qualifiers
     source          1..7509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    64..405
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             472..888
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     818..840
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     856..872
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1513..1525)
                     /gene="Pichia pastoris ADE2"
                     /label=PpADE2 HC promoter
                     /note="weak 13-bp promoter fragment that requires high-copy
                     integration for efficient expression"
     misc_feature    1985..2552
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2002..2554
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2146..2163)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2373..2392
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2566..2568,2569..2571,2572..3285)
                     /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(2611..2632)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2872..2891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3219..3240
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3387..3901
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4063..4079
                     /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(4087..4093,4094..4111,4112..4117))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4132..4153
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4270..4287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4441..5029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4521..4540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6233..6251
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6289..6311)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6411..6430
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6624..6646
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6774..6793
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6993..7509
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga
       61 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
      121 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
      181 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
      241 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
      301 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
      361 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
      421 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
      481 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
      541 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
      601 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
      661 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
      721 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
      781 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
      841 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattagatct
      901 ctcgagatgg actacaaaga cgatgacgac aagatgaaca tacacaggtc tacccctatc
      961 acaatagcga gatatgggag atcgcggaac aaaacccagg atttcgaaga gctgtcgtct
     1021 ataaggtccg ctgagcccag ccagagtttc agcccgaacc ttggctctcc gagcccgccc
     1081 gagactccga acttgtcgca ttgcgtttct tgcatcggga aatacttact gttggagcct
     1141 ctggagggag accacgtttt ccgcgctgtg catctgcaca gcggagagga gctggtttgc
     1201 aaggtgtttg agatcagctg ctaccaggag tccctggccc cctgcttctg cctgtctgcc
     1261 catagcaaca tcaaccaaat cacggaaatc ctcctgggag agaccaaagc ctatgtgttc
     1321 tttgagcgaa gctatggaga catgcattcc tttgtccgca cttgtaagaa gctgagggag
     1381 gaggaggcag cccgactgtt ctaccagatt gcctcagctg tggcccattg ccacgatgga
     1441 ggcctggtgc tgcgtgacct caagctgcgg aaatttatct tcaaggatga agagaggact
     1501 cgtgtcaagc tggagagttt ggaagacgct tacattctcc ggggtgatga tgactcactc
     1561 tctgacaagc atggctgccc agcgtatgtc agcccagaga tcttgaacac cagcggcagt
     1621 tattcgggca aggcagcgga cgtgtggagc ctgggggtaa tgctgtacac catgttggtg
     1681 gggcgttacc ctttccatga cattgagcct agttctctct tcagtaagat ccgcaggggc
     1741 cagttcaaca ttccagaaac tctgtctccc aaggccaagt gcctcatccg aagcatcctg
     1801 cgacgggagc cgtcagagcg gctgacctcg caggaaattc tggaccatcc ttggttttct
     1861 acagatttta gtgtctcaaa ttcgggattt ggtgctaaag aggcgtgtga ccagctggtg
     1921 ccagacgtca acatggagga gaacttggac cctttcttta actgaggatt cggaaagggc
     1981 gaattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2041 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2101 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2161 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2221 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2281 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2341 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2401 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     2461 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     2521 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc
     2581 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2641 cacaagttca gcgtgtctgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2701 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2761 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2821 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2881 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2941 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3001 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggcgaac
     3061 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     3121 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     3181 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3241 accgccgccg ggatcactct cggcatggac gagctgtaca agtaatgaat taattaagaa
     3301 ttatcaagct tatcgatacc gtcgacctgc agccaagctt atcgataaaa taaaagattt
     3361 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     3421 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     3481 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     3541 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3601 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     3661 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3721 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     3781 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     3841 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     3901 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
     3961 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
     4021 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     4081 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4141 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4201 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     6241 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6301 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6361 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     6421 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     6481 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6541 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6601 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     6661 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     6721 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     6781 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     6841 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     6901 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     6961 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     7021 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     7081 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     7141 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     7201 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     7261 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     7321 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     7381 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     7441 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     7501 tctttcatt
//
