LOCUS       sequence-489523-        7312 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for a knockdown of MYB in human Megakaryocyte/Erythroid 
            Progenitors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7312)
  AUTHORS   Sanada C, Xavier-Ferrucio J, Lu YC, Min E, Zhang PX, Zou S, Kang E, 
            Zhang M, Zerafati G, Gallagher PG, Krause DS
  TITLE     Adult human megakaryocyte-erythroid progenitors are in the 
            CD34+CD38mid fraction.
  JOURNAL   Blood. 2016 Aug 18;128(7):923-33. doi: 10.1182/blood-2016-01-693705.
            Epub 2016 Jun 6.
  PUBMED    27268089
REFERENCE   2  (bases 1 to 7312)
  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-2016-01-693705"; journalName: "Blood"; date: 
            "2016-08-18- 18"; volume: "128"; issue: "7"; pages: "923-33"
COMMENT     Sequence Label: shMYB-2
FEATURES             Location/Qualifiers
     source          1..7312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2134..2367
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2552..2596
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2985..3233
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2985..3005
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3156..3175
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    3372..3489
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3538..4048
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3967..3985
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      4059..4068
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4065..4067,4068..4070,4071..4781)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4065..4067,4068..4070,4071..4781)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4110..4131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4371..4390)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4718..4739
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4888..5121
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5193..5314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5230..5249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5284..5303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5354..5489
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5509..5528)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5510..5528)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5538..5555)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5538..5554)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5547..5569)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5696..6151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5783..5802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5993..6014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6177..6281
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6282..6350,6351..7142)
                     /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(6500..6519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     2041 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     2101 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     2161 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     2221 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     2281 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     2341 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     2401 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     2461 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     2521 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     2581 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     2641 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     2701 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     2761 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
     2821 atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc ctcgagcggc
     2881 ccggggctag ctgaggatag cagagaagga cgcgtgatcc tgcagctcct gttctcacct
     2941 tcgcggccgc tctagagtcg actacgtagg ccgccccctt caccgagggc ctatttccca
     3001 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     3061 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     3121 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     3181 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggaaca
     3241 gaatggaaca gatgacctcg aggtcatctg ttccattctg ttctttttga attctacgta
     3301 gatatctgat caccatacgt atggcccggg taattctcga cctcgagaca aatggcagta
     3361 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     3421 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     3481 caaaattttc gggtttatta cagggacagc agagatccac tttggccgcg gctcgagggg
     3541 gttggggttg cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg
     3601 gcgtggttcc gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt
     3661 tcgcagcgtc acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc
     3721 tccgccccta agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga
     3781 agccgcacgt ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg
     3841 ccgaccgcga tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg
     3901 ccgggaaggg gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc
     3961 cgcgcggtgt tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt
     4021 tgaccgaatc accgacctct ctccccaggg ggatcctcgc caccatggtg agcaagggcg
     4081 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4141 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4201 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     4261 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     4321 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4381 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4441 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4501 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4561 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4621 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4681 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4741 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc agatctcgac
     4801 tagctagtag ctagctggta cctttaagac caatgactta caaggcagct gtagatctta
     4861 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     4921 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     4981 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5041 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5101 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5161 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     5221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     5341 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5401 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5461 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
     5521 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5581 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5641 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     5701 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5761 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5821 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5881 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5941 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6001 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6061 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6121 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     6181 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6241 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6301 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6361 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6421 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6481 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6541 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6601 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6661 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6721 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6781 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6841 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6901 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6961 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     7021 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     7081 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     7141 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     7201 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     7261 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
