LOCUS       sequence-319146-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid of mutant MYB in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  AUTHORS   Weng H, Huang H, Wu H, Qin X, Zhao BS, Dong L, Shi H, Skibbe J, Shen
            C, Hu C, Sheng Y, Wang Y, Wunderlich M, Zhang B, Dore LC, Su R, Deng
            X, Ferchen K, Li C, Sun M, Lu Z, Jiang X, Marcucci G, Mulloy JC, 
            Yang J, Qian Z, Wei M, He C, Chen J
  TITLE     METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and 
            Promotes Leukemogenesis via mRNA m(6)A Modification.
  JOURNAL   Cell Stem Cell. 2018 Feb 1;22(2):191-205.e9. doi: 
            10.1016/j.stem.2017.11.016. Epub 2017 Dec 28.
  PUBMED    29290617
REFERENCE   2  (bases 1 to 6824)
  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.stem.2017.11.016"; journalName: "Cell Stem Cell"; date: 
            "2018-02-1- 1"; volume: "22"; issue: "2"; pages: "191-205.e9"
COMMENT     Sequence Label: pmiR-report-MYB-mut
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        98..401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        402..605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     556..576
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             630..2279
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(696..714)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2789..2806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2789..2805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2820)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3014..3033)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3195..3511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3362..3497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3608..4207
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3608..3627)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4104..4124
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4437..4518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4474..4493)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4606..4710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4711..4779,4780..5571)
                     /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     complement(4929..4948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5742..6330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6231..6250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6484..6501
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6618..6639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6654..6659,6660..6677,6678..6684)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6692..6708
                     /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)."
     primer_bind     6697..6719
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6716..6732
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6716..6732
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6752..6770
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6752..6769
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6782..6801
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6782..6800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaagggagga gaagcatgaa ttcaaggtac cagatcttag ttattaatag taatcaatta
       61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      601 gagctggttt agtgaaccgt ggatccacca tggaagacgc caaaaacata aagaaaggcc
      661 cggcgccatt ctatcctcta gaggatggaa ccgctggaga gcaactgcat aaggctatga
      721 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtgaaca
      781 tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
      841 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
      901 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
      961 aacgtgaatt gctcaacagt atgaacattt cgcagcctac cgtagtgttt gtttccaaaa
     1021 aggggttgca aaaaattttg aacgtgcaaa aaaaattacc aataatccag aaaattatta
     1081 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
     1141 atctacctcc cggttttaat gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa
     1201 caattgcact gataatgaat tcctctggat ctactgggtt acctaagggt gtggcccttc
     1261 cgcatagaac tgcctgcgtc agattctcgc atgccagaga tcctattttt ggcaatcaaa
     1321 tcgttccgga tactgcgatt ttaagtgttg tcccattcca tcacggtttt ggaatgttta
     1381 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
     1441 agctgttttt acgatccctt caggattaca aaattcaaag tgcgttgcta gtaccaaccc
     1501 tattttcatt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
     1561 aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct
     1621 tccatcttcc agggatacga caaggatatg ggctcactga gactacatca gctattctga
     1681 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
     1741 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcagaga ggcgaattat
     1801 gtgtcagagg acctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
     1861 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
     1921 acttcttcat agttgaccgc ttgaagtctt taattaaata caaaggatat caggtggccc
     1981 ccgctgaatt ggaatcgata ttgttacaac accccaacat cttcgacgcg ggcgtggcag
     2041 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
     2101 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
     2161 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
     2221 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag tccaaattgc
     2281 tcgagggtgg aatctccaac tgataaatca ggaatcttct tctgctcaca ccactgggaa
     2341 ggggtcagtc tgaataccca actgttcacg cagacctcgc ctgtggcaga tgcaccgaat
     2401 attcttacaa gctccgtttt aatggcacca gcatcagaag atgaagtcaa tgttctcaaa
     2461 gcatttacag tacctaaaaa caggtccctg gcgagcccct tgcagccttg tagcagtacc
     2521 tgggaacctg catcctgtgg aaagatggag gagcagatga catcttccag tcaagctcgt
     2581 aaatacgtga atgcattctc agcccggtcg ctggtcatgt gagtcatttc cagctcgagt
     2641 gatgaaagct gcgcactagt gagggagctc ataggccggc atagacgcgt gcgtgatatc
     2701 ataggtttaa acagttaagc ttaataaagg atcttttatt ttcattggat ctgtgtgttg
     2761 gttttttgta tgcggccgct agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     2821 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2881 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2941 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3001 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3061 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3121 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3181 cgagacgaaa gattggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3241 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3301 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3361 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3421 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3481 aggctttttt ggaggcctag gcttttgcaa aaagctagct tgcatgcctg caggtcggcc
     3541 gccacgaccg gtgccgccac catcccctga cccacgcccc tgacccctca caaggagacg
     3601 accttccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     3661 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3721 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3781 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     3841 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3901 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     3961 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     4021 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     4081 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     4141 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     4201 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg
     4261 ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc gaggcccacc
     4321 gactctagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     4381 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     4441 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4501 aagcattttt ttcactgcaa tctcgtgata cgcctatttt tataggttaa tgtcatgata
     4561 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4681 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4741 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4801 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4861 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4921 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4981 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5041 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5101 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5161 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5221 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5281 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5341 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5401 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5461 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5521 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttttttgccg
     5821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5881 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6421 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6481 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6541 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6601 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6661 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6721 aacagctatg acatgattac gaattgcaac gatttaggtg acactataga agagaaggaa
     6781 ttaatacgac tcactatagg gagagagaga gaattaccct cact
//
