LOCUS       sequence-302048-        6824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid of WT 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-WT
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..350
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        351..554
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     505..525
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             579..2228
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(645..663)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2738..2755)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2738..2754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2747..2769)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2963..2982)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3144..3460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3311..3446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3557..4156
                     /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(3557..3576)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4053..4073
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4386..4467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4423..4442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4555..4659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4660..4728,4729..5520)
                     /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(4878..4897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5691..6279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6180..6199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6433..6450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6567..6588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6603..6608,6609..6626,6627..6633)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6641..6657
                     /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     6646..6668
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6665..6681
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6665..6681
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6701..6719
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6701..6718
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6731..6750
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6731..6749
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
       61 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      121 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      181 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      241 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      301 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      361 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      421 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      481 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      541 atataagcag agctggttta gtgaaccgtg gatccaccat ggaagacgcc aaaaacataa
      601 agaaaggccc ggcgccattc tatcctctag aggatggaac cgctggagag caactgcata
      661 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
      721 aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt tcggttggca gaagctatga
      781 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
      841 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
      901 tttataatga acgtgaattg ctcaacagta tgaacatttc gcagcctacc gtagtgtttg
      961 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaaattacca ataatccaga
     1021 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1081 tcacatctca tctacctccc ggttttaatg aatacgattt tgtaccagag tcctttgatc
     1141 gtgacaaaac aattgcactg ataatgaatt cctctggatc tactgggtta cctaagggtg
     1201 tggcccttcc gcatagaact gcctgcgtca gattctcgca tgccagagat cctatttttg
     1261 gcaatcaaat cgttccggat actgcgattt taagtgttgt cccattccat cacggttttg
     1321 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     1381 ttgaagaaga gctgttttta cgatcccttc aggattacaa aattcaaagt gcgttgctag
     1441 taccaaccct attttcattc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     1501 atttacacga aattgcttct gggggcgcac ctctttcgaa agaagtcggg gaagcggttg
     1561 caaaacgctt ccatcttcca gggatacgac aaggatatgg gctcactgag actacatcag
     1621 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     1681 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcagagag
     1741 gcgaattatg tgtcagagga cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     1801 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     1861 aagacgaaca cttcttcata gttgaccgct tgaagtcttt aattaaatac aaaggatatc
     1921 aggtggcccc cgctgaattg gaatcgatat tgttacaaca ccccaacatc ttcgacgcgg
     1981 gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2041 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2101 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     2161 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaagt
     2221 ccaaattgct cgagggtgga atctccaact gataaatcag gaaacttctt ctgctcacac
     2281 cactgggaag gggacagtct gaatacccaa ctgttcacgc agacctcgcc tgtggcagat
     2341 gcaccgaata ttcttacaag ctccgtttta atggcaccag catcagaaga tgaagacaat
     2401 gttctcaaag catttacagt acctaaaaac aggtccctgg cgagcccctt gcagccttgt
     2461 agcagtacct gggaacctgc atcctgtgga aagatggagg agcagatgac atcttccagt
     2521 caagctcgta aatacgtgaa tgcattctca gcccggacgc tggtcatgtg agacatttcc
     2581 agctcgagtg atgaaagctg cgcactagtg agggagctca taggccggca tagacgcgtg
     2641 cgtgatatca taggtttaaa cagttaagct taataaagga tcttttattt tcattggatc
     2701 tgtgtgttgg ttttttgtat gcggccgcta gcttggcact ggccgtcgtt ttacaacgtc
     2761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2881 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     2941 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     3001 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     3061 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     3121 cgaaacgcgc gagacgaaag attggtgtgg aaagtcccca ggctccccag caggcagaag
     3181 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3301 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctagctt gcatgcctgc
     3481 aggtcggccg ccacgaccgg tgccgccacc atcccctgac ccacgcccct gacccctcac
     3541 aaggagacga ccttccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3601 cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3661 caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3721 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3781 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3841 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3901 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3961 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     4021 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4081 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4141 caagcccggt gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg
     4201 accccatggc tccgaccgaa gccacccggg gcggccccgc cgaccccgca cccgcccccg
     4261 aggcccaccg actctagagg atcataatca gccataccac atttgtagag gttttacttg
     4321 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4381 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4441 tcacaaataa agcatttttt tcactgcaat ctcgtgatac gcctattttt ataggttaat
     4501 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4561 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4861 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5521 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5761 tttttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5821 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6361 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6421 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6481 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6541 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6601 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     6661 cacacaggaa acagctatga catgattacg aattgcaacg atttaggtga cactatagaa
     6721 gagaaggaat taatacgact cactataggg agagagagag aattaccctc actaaaggga
     6781 ggagaagcat gaattcaagg taccagatct tagttattaa tagt
//
