LOCUS       sequence-252744-        7189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector of GFP-tagged human BMI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7189)
  AUTHORS   Hossain S, Iwasa H, Sarkar A, Maruyama J, Arimoto-Matsuzaki K, Hata 
            Y
  TITLE     The RASSF6 Tumor Suppressor Protein Regulates Apoptosis and Cell 
            Cycle Progression via Retinoblastoma Protein.
  JOURNAL   Mol Cell Biol. 2018 Aug 15;38(17). pii: MCB.00046-18. doi: 
            10.1128/MCB.00046-18. Print 2018 Sep 1.
  PUBMED    29891515
REFERENCE   2  (bases 1 to 7189)
  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"; journalName: "Mol Cell 
            Biol."; date: "2018-08-15"; volume: "38(17). pii: MCB.00046-18. 
            doi"; pages: " 10.1128/MCB.00046-18. Print 2018 Sep 1"
COMMENT     Sequence Label: pCIneoGFP-BMI1
FEATURES             Location/Qualifiers
     source          1..7189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          373..505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     488..507
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      584..593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(590..592,593..595,596..1306)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(590..592,593..595,596..1306)
                     /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(635..656)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(896..915)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1243..1264
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2376..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2683..3138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2770..2789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2980..3001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3255..3612
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3255..3446
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3278..3298)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3463..3598
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3663..4457
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3717..3736)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4327..4346
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4521..4569
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4610..4629)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4729..4751
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4789..4807)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4875..4979
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4980..5048,5049..5840)
                     /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(5198..5217)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6011..6599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6500..6519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6810..7189
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
      241 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
      301 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
      361 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      421 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      481 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      541 tagagtactt aatacgactc actataggct agcgctaccg gtcgccacca tggtgagcaa
      601 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      661 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      721 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      781 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      841 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      901 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      961 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1021 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     1081 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1141 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1201 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1261 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc
     1321 tcgagctcaa gcttcgacgc gtatgcatcg aacaacgaga atcaagatca ctgagctaaa
     1381 tccccacctg atgtgtgtgc tttgtggagg gtacttcatt gatgccacaa ccataataga
     1441 atgtctacat tccttctgta aaacgtgtat tgttcgttac ctggagacca gcaagtattg
     1501 tcctatttgt gatgtccaag ttcacaagac cagaccatta ctgaatataa ggtcagataa
     1561 aactctccaa gatattgtat acaaattagt tccagggctt ttcaaaaatg aaatgaagag
     1621 aagaagggat ttttatgcag ctcatccttc tgctgatgct gccaatggct ctaatgaaga
     1681 tagaggagag gttgcagatg aagataagag aattataact gatgatgaga taataagctt
     1741 atccattgaa ttctttgacc agaacagatt ggatcggaaa gtaaacaaag acaaagagaa
     1801 atctaaggag gaggtgaatg ataaaagata cttacgatgc ccagcagcaa tgactgtgat
     1861 gcacttaaga aagtttctca gaagtaaaat ggacatacct aatactttcc agattgatgt
     1921 catgtatgag gaggaacctt taaaggatta ttatacacta atggatattg cctacattta
     1981 tacctggaga aggaatggtc cacttccatt gaaatacaga gttcgaccta cttgtaaaag
     2041 aatgaagatc agtcaccaga gagatggact gacaaatgct ggagaactgg aaagtgactc
     2101 tgggagtgac aaggccaaca gcccagcagg aggtattccc tccacctctt cttgtttgcc
     2161 tagccccagt actccagtgc agtctcctca tccacagttt cctcacattt ccagtactat
     2221 gaatggaacc agcaacagcc ccagcggtaa ccaccaatct tcttttgcca atagacctcg
     2281 aaaatcatca gtaaatgggt caccagcaac ttcttctggt tgagtcgacg cggccgcttc
     2341 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
     2401 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     2581 atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc
     2641 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     2701 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2761 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2821 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2881 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2941 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3001 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3061 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3121 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3181 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3241 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3361 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3541 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3601 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     3661 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3721 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3781 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3841 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3901 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3961 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4021 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4081 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4141 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4201 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4261 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4321 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4381 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4441 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4501 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     4561 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
     4621 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4681 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4741 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4801 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4861 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4921 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4981 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5041 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5101 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5161 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5221 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5281 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5341 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5401 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5461 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5521 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5581 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5641 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5701 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5761 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5821 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5881 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5941 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6001 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6061 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     6661 ctcgacagat cttcaatatt ggccattagc catattattc attggttata tagcataaat
     6721 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     6781 tggctcatgt ccaatatgac cgccatgttg gcattgatta ttgactagtt attaatagta
     6841 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6901 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6961 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7021 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     7081 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     7141 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
