LOCUS       sequence-311510-        6893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express human EMI1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  AUTHORS   Marzio A, Puccini J, Kwon Y, Maverakis NK, Arbini A, Sung P, 
            Bar-Sagi D, Pagano M
  TITLE     The F-Box Domain-Dependent Activity of EMI1 Regulates PARPi 
            Sensitivity in Triple-Negative Breast Cancers.
  JOURNAL   Mol Cell. 2019 Jan 17;73(2):224-237.e6. doi: 
            10.1016/j.molcel.2018.11.003. Epub 2018 Dec 13.
  PUBMED    30554948
REFERENCE   2  (bases 1 to 6893)
  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.molcel.2018.11.003"; journalName: "Mol Cell"; date: 
            "2019-01-17- 17"; volume: "73"; issue: "2"; pages: "224-237.e6"
COMMENT     Sequence Label: pcDNA3.1 2xFLAG-2xSTREP EMI1
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..946
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             962..985
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1001..1024
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1061..1084
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2484..2501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2490..2714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2760..3188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2847..2866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3057..3078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3197..3217)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3202..3531
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3382..3517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3444..3463
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3598..4392
                     /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(3652..3671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4262..4281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4566..4687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4603..4622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4657..4676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4736..4752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4736..4752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4749..4771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4760..4776
                     /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(4784..4790,4791..4808,4809..4814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4829..4850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4967..4984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5138..5726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5218..5237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5897..6688,6689..6757))
                     /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     6520..6539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6758..6862)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttacca tggactacaa ggacgacgat gacaaggtac caggatctat
      961 ggattataaa gatgatgatg ataaagggtc ggccgccagc tggagccacc ctcagttcga
     1021 gaagggagga ggaagcggcg gaggcagcgg aggaggaagc tggagccacc cgcagttcga
     1081 gaaaggagct ggatctaata tgagccggcg cccctgcagc tgcgccctac ggccaccccg
     1141 ctgctcctgc agcgccagcc ccagcgcagt gacagccgcc gggcgccctc gaccctcgga
     1201 tagttgtaaa gaagaaagtt ctaccctttc tgtcaaaatg aagtgtgatt ttaattgtaa
     1261 ccatgttcat tccggactta aactggtaaa acctgatgac attggaagac tagtttccta
     1321 cacccctgca tatttggaag gttcctgtaa agactgcatt aaagactatg aaaggctgtc
     1381 atgtattggg tcaccgattg tgagccctag gattgtagaa cttgaaactg aaagcaagcg
     1441 cttgcataac aaggaaaatc aacatgtgca acagacactt aatagtacaa atgaaataga
     1501 agcactagag accagtagac tttatgaaga cagtggctat tcctcatttt ctctacaaag
     1561 tggcctcagt gaacatgaag aaggtagcct cctggaggag aatttcggtg acagtctaca
     1621 atcctgcctg ctacaaatac aaagcccaga ccaatatccc aacaaaaact tgctgccagt
     1681 tcttcatttt gaaaaagtgg tttgttcaac attaaaaaag aatgcaaaac gaaatcctaa
     1741 agtagatcgg gagatgctga aggaaattat agccagagga aattttagac tgcagaatat
     1801 aattggcaga aaaatgggcc tagaatgtgt agatattctc agcgaactct ttcgaagggg
     1861 actcagacat gtcttagcaa ctattttagc acaactcagt gacatggact taatcaatgt
     1921 gtctaaagtg agcacaactt ggaagaagat cctagaagat gataaggggg cattccagtt
     1981 gtacagtaaa gcaatacaaa gagttaccga aaacaacaat aaattttcac cccatgcttc
     2041 aaccagagaa tatgttatgt tcagaacccc actggcttct gttcagaaat cagcagccca
     2101 gacttctctc aaaaaagatg ctcaaaccaa gttatccaat caaggtgatc agaaaggttc
     2161 tacttatagt cgacacaatg aattctctga ggttgccaag acattgaaaa agaacgaaag
     2221 cctcaaagcc tgtattcgct gtaattcacc tgcaaaatat gattgctatt tacaacgggc
     2281 aacctgcaaa cgagaaggct gtggatttga ttattgtacg aagtgtctct gtaattatca
     2341 tactactaaa gactgttcag atggcaagct cctcaaagcc agttgtaaaa taggtcccct
     2401 gcctggtaca aagaaaagca aaaagaattt acgaagattg tgatccctcg agtctagagg
     2461 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2521 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2581 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2641 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2701 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2761 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2821 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2881 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2941 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3001 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3061 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3121 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3181 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3361 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3421 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3481 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3541 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4561 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4621 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4681 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4741 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4801 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4861 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4921 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4981 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5041 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5101 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5161 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5221 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5281 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5341 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5401 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5461 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5521 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5581 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5641 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5701 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5761 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5821 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5881 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5941 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6001 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6061 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6121 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6181 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6241 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6301 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6361 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6421 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6481 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6541 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6601 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6661 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6721 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6781 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6841 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
