LOCUS       sequence-203810-        6544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retrovirally express RPA3 shRNA with GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  AUTHORS   Xu Y, Milazzo JP, Somerville TDD, Tarumoto Y, Huang YH, Ostrander 
            EL, Wilkinson JE, Challen GA, Vakoc CR
  TITLE     A TFIID-SAGA Perturbation that Targets MYB and Suppresses Acute 
            Myeloid Leukemia.
  JOURNAL   Cancer Cell. 2018 Jan 8;33(1):13-28.e8. doi: 
            10.1016/j.ccell.2017.12.002.
  PUBMED    29316427
REFERENCE   2  (bases 1 to 6544)
  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.ccell.2017.12"; journalName: "Cancer Cell"; date: 
            "2018-01-8- 8"; volume: "33"; issue: "1"; pages: "13-28.e8"
COMMENT     Sequence Label: MLSE-shRPA3.457
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     ncRNA           1592..1606
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(1804..1824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1809..2138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1989..2124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(2145..2147,2148..2150,2151..2864)
                     /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(2190..2211)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2451..2470)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2798..2819
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2937..3451
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3613..3629
                     /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(3637..3643,3644..3661,3662..3667))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3682..3703
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3820..3837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3991..4579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4071..4090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4750..5541,5542..5610))
                     /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     5373..5392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5611..5715)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5783..5801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5839..5861)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5961..5980
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6174..6196
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6324..6343
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6543..6544,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctctcga ctagggataa cagggtaatt
     1441 gtttgaatga ggcttcagta ctttacagaa tcgttgcctg cacatcttgg aaacacttgc
     1501 tgggattact tcgacttctt aacccaacag aaggctcgag aaggtatatt gctgttgaca
     1561 gtgagcgcgc gactcctata atttctaatt agtgaagcca cagatgtaat tagaaattat
     1621 aggagtcgct tgcctactgc ctcggacttc aaggggctag aattcgagca attatcttgt
     1681 ttactaaaac tgaatacctt gctatctctt tgatacattt ttacaaagct gaattaaaat
     1741 ggtataaatt aaatcacttt tttcaattga cgcgtaattc gccagcacag tggtcgaccc
     1801 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     1861 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     1981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2041 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2101 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcatggtg agcaagggcg
     2161 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2221 acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2281 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2341 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2401 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2461 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2521 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2581 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact
     2641 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2701 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2761 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2821 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaatgaatt aattaagaat
     2881 tatcaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     2941 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3001 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     3061 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     3121 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     3181 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     3241 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     3301 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     3361 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     3421 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     3481 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     3541 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     3601 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3661 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     3721 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     3781 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5761 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     5821 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5881 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5941 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6001 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     6061 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6121 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     6181 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6241 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6301 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6361 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     6421 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     6481 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     6541 ggaa
//
