LOCUS       sequence-255694-        5595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To investigate activity of a 800 bp region upstream of mouse Trib2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5595)
  AUTHORS   Rishi L, Hannon M, Salome M, Hasemann M, Frank AK, Campos J, Timoney
            J, O'Connor C, Cahill MR, Porse B, Keeshan K
  TITLE     Regulation of Trib2 by an E2F1-C/EBPalpha feedback loop in AML cell 
            proliferation.
  JOURNAL   Blood. 2014 Apr 10;123(15):2389-400. doi: 
            10.1182/blood-2013-07-511683. Epub 2014 Feb 10.
  PUBMED    24516045
REFERENCE   2  (bases 1 to 5595)
  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-2013-07-511683"; journalName: "Blood"; date: 
            "2014-04-10- 10"; volume: "123"; issue: "15"; pages: "2389-400"
COMMENT     Sequence Label: pGL3 T2P 800
FEATURES             Location/Qualifiers
     source          1..5595
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..97
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     46..65
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      963..972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             969..2621
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1035..1053)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2662..2783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2673..2692)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2727..2746
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3031..3048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3202..3790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3282..3301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3961..4752,4753..4821))
                     /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     4584..4603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4822..4926)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4953..5408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5040..5059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5250..5271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5539..5587
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
       61 gtccccagtg caagtgcagg tgccagaaca tttctctatc gataggtacc tgtgcctgcc
      121 ctactccgcg cactccctac tcagcttttg caatcttttg ttgccactgc tgaccgcccc
      181 aagttaaaag cgcctttgct attttttttt tctgtctttt tttttttttc ttgagagaga
      241 cagctctgcg attttttgga aaagcctcag acgccatcta cagttaagac ttaggtaact
      301 gccctctcta gcacccccct tacacgcccc ccaccctttc caccaaaaag aggggggtgc
      361 agcgcggatt ctggctgccg cgcggcgtga gccggtagac ccgagcttat ttcctttttc
      421 tttttgtttg gtttctaacg cgtggagggc gagccggcgc cgcgcgctcc ctgaagactg
      481 cacaaactcc acgcagggct tctccgcccg gtctgcggat cctcagctgg ggatcgctca
      541 gaagcccggc gctgcagctc ctcaccccag aggcacgctc actcgtccag atccacgctg
      601 cgaacagaga cccactgagt ccagcgtgcg gttctgcacc gcgctggcag cttctgggta
      661 acaaaaggac ccgagttgtc cgcagagcga gcacccccgg gagcggggct cgcagccggg
      721 gaccagccct gcagcgccca tctgggggct agttcttaac tcttcctcca cggagcccca
      781 gacgggtccc ctcccttctt gatcctttta agtcggtagc accgaggcgc ctgcaccggc
      841 gcggctcatc catctctcca gaggggtttt ttggtttgtt tgtttgtttt gtcgtgtgcg
      901 atcctcacac tcctcgagat ctgcgatcta agtaagcttg gcattccggt actgttggta
      961 aagccaccat ggaagacgcc aaaaacataa agaaaggccc ggcgccattc tatccgctgg
     1021 aagatggaac cgctggagag caactgcata aggctatgaa gagatacgcc ctggttcctg
     1081 gaacaattgc ttttacagat gcacatatcg aggtggacat cacttacgct gagtacttcg
     1141 aaatgtccgt tcggttggca gaagctatga aacgatatgg gctgaataca aatcacagaa
     1201 tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc ggtgttgggc gcgttattta
     1261 tcggagttgc agttgcgccc gcgaacgaca tttataatga acgtgaattg ctcaacagta
     1321 tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa ggggttgcaa aaaattttga
     1381 acgtgcaaaa aaagctccca atcatccaaa aaattattat catggattct aaaacggatt
     1441 accagggatt tcagtcgatg tacacgttcg tcacatctca tctacctccc ggttttaatg
     1501 aatacgattt tgtgccagag tccttcgata gggacaagac aattgcactg atcatgaact
     1561 cctctggatc tactggtctg cctaaaggtg tcgctctgcc tcatagaact gcctgcgtga
     1621 gattctcgca tgccagagat cctatttttg gcaatcaaat cattccggat actgcgattt
     1681 taagtgttgt tccattccat cacggttttg gaatgtttac tacactcgga tatttgatat
     1741 gtggatttcg agtcgtctta atgtatagat ttgaagaaga gctgtttctg aggagccttc
     1801 aggattacaa gattcaaagt gcgctgctgg tgccaaccct attctccttc ttcgccaaaa
     1861 gcactctgat tgacaaatac gatttatcta atttacacga aattgcttct ggtggcgctc
     1921 ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt ccatctgcca ggtatcaggc
     1981 aaggatatgg gctcactgag actacatcag ctattctgat tacacccgag ggggatgata
     2041 aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc gaaggttgtg gatctggata
     2101 ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg tgtgagaggt cctatgatta
     2161 tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt gattgacaag gatggatggc
     2221 tacattctgg agacatagct tactgggacg aagacgaaca cttcttcatc gttgaccgcc
     2281 tgaagtctct gattaagtac aaaggctatc aggtggctcc cgctgaattg gaatccatct
     2341 tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg tcttcccgac gatgacgccg
     2401 gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa gacgatgacg gaaaaagaga
     2461 tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa gttgcgcgga ggagttgtgt
     2521 ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga cgcaagaaaa atcagagaga
     2581 tcctcataaa ggccaagaag ggcggaaaga tcgccgtgta attctagagt cggggcggcc
     2641 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2701 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2761 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     2821 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     2881 ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     2941 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     3001 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3061 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3121 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3181 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3241 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3301 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3361 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3421 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3481 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3541 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3601 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3661 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3721 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3781 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3841 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3901 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3961 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4021 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4081 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4141 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4201 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4261 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4321 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4381 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4441 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4501 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4561 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4621 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4681 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4741 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4801 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4921 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
     4981 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5041 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5101 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5161 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5221 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5281 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5341 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5401 tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5461 ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga
     5521 ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     5581 ttgtgtgaat cgata
//
