LOCUS       sequence-402527-        8594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MDM4 overexpression vector with blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8594)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 8594)
  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: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: pLVX-Bsd-CMV-MDM4
FEATURES             Location/Qualifiers
     source          1..8594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    75..99
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1606..1630)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1642..1662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1647..1976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1827..1962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1889..1908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2038..2433
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2462..3050
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2515..2535)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2933..2944)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3256..3489
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(3606..3622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3606..3622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3619..3641)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3630..3646
                     /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(3654..3660,3661..3678,3679..3684))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3699..3720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3837..3854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4008..4596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4088..4107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4767..5558,5559..5627))
                     /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     5390..5409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5628..5732)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    5780..5914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5817..5836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5871..5890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        5997..6015
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6043..6269
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6270..6450
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6497..6622
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7123..7356
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7541..7585
                     /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"
     misc_feature    7848..7963
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     primer_bind     8002..8021
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8002..8020
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        8118..8421
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(8423..8594,1..31)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(8576..8594,1..2)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 tgtacggtgg gaggtctata taagcagacg tcgtttagtg aaccgtcaga tcactagatg
       61 ctttattgga tctaacaagt ttgtacaaaa aagcaggctt ggaaggagtt cgaaccatga
      121 catcattttc cacctctgct cagtgttcaa catctgacag tgcttgcagg atctctcctg
      181 gacaaatcaa tcaggtacga ccaaaactgc cgcttttgaa gattttgcat gcagcaggtg
      241 cgcaaggtga aatgttcact gttaaagagg tcatgcacta tttaggtcag tacataatgg
      301 tgaagcaact ttatgatcag caggagcagc atatggtata ttgtggtgga gatcttttgg
      361 gagaactact gggacgtcag agcttctccg tgaaagaccc aagccctctc tatgatatgc
      421 taagaaagaa tcttgtcact ttagccactg ctactacaga tgctgctcag actctcgctc
      481 tcgcacagga tcacagtatg gatattccaa gtcaagacca actgaagcaa agtgcagagg
      541 aaagttccac ttccagaaaa agaactacag aagacgatat ccccacactg cctacctcag
      601 agcataaatg catacattct agagaagatg aagacttaat tgaaaattta gcccaagatg
      661 aaacatctag gctggacctt ggatttgagg agtgggatgt agctggcctg ccttggtggt
      721 ttttaggaaa cttgagaagc aactatacac ctagaagtaa tggctcaact gatttacaga
      781 caaatcagga tgtgggtact gccattgttt cagatactac agatgacttg tggtttttga
      841 atgagtcagt atcagagcag ttaggtgttg gaataaaagt tgaagctgct gatactgaac
      901 aaacaagtga agaagtaggg aaagtaagtg acaaaaaggt gattgaagtg ggaaaaaatg
      961 atgacctgga ggactctaag tccttaagtg atgataccga tgtagaggtt acctctgagg
     1021 atgagtggca gtgtactgaa tgcaagaaat ttaactctcc aagcaagagg tactgttttc
     1081 gttgttgggc cttgaggaag gattggtatt cagattgttc aaagttaacc cattctctct
     1141 ccacgtctga tatcactgcc atacctgaaa aggaaaatga aggaaatgat gtccctgatt
     1201 gtcgaagaac catttcggct cctgtcgtta gacctaaaga tgcgtatata aagaaagaaa
     1261 actccaaact ttttgatccc tgcaactcag tggaattctt ggatttggct cacagttctg
     1321 aaagccaaga gaccatctca agcatgggag aacagttaga taacctttct gaacagagaa
     1381 cagatacaga aaacatggag gattgccaga atctcttgaa gccatgtagc ttatgtgaga
     1441 aaagaccacg agacgggaac attattcatg gaaggacggg ccatcttgtc acttgttttc
     1501 actgtgccag aagactaaag aaggctgggg cttcatgccc tatttgcaag aaagagattc
     1561 agctggttat taaggttttt atagcatagc tcgagtgcgg ccgcaaccca gctttcttgt
     1621 acaaagtggt tctcgagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     1741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1801 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     1861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     1921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     1981 tcccgggagc ttgtatatcc attttcggat ctgatcggcg cgccataccg gtccaccatg
     2041 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     2101 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     2161 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     2221 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     2281 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     2341 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     2401 gaattgctgc cctctggtta tgtgtgggag ggcctgcagc tgcagtagac gcgtctggaa
     2461 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2521 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2581 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2641 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2701 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2761 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2821 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2881 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2941 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3001 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     3061 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     3121 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     3181 cctcaggacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     3241 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     3301 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     3361 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     3421 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     3481 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     3541 gaatatcaga gagtgagagg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa
     3601 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3661 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3721 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3781 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3841 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3901 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3961 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4021 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4081 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4141 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4201 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4261 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4321 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4381 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4441 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4501 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4561 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4621 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4681 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4741 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4801 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4861 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4921 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4981 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5041 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5101 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5161 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5221 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5281 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5341 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5401 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5461 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5521 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5581 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5641 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5701 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5761 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5881 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa
     5941 tcatcccaaa cttcccaccc cataccctat taccactgcc aattaccctg tggcgcaatt
     6001 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     6061 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     6121 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     6181 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     6241 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     6301 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6361 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6421 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     6481 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     6541 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     6601 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     6661 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     6721 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     6781 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     6841 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     6901 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     6961 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaatta attggagaag
     7021 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7081 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7141 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     7201 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     7261 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     7321 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     7381 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     7441 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     7501 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     7561 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     7621 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     7681 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     7741 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     7801 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     7861 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     7921 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     7981 acagcagaga tccagtttat ctaatacgac tcactatagg gagagagaga gaattaccct
     8041 cactaaaggg aggagaagca tgaattctag taatcaatta cggggtcatt agttcatagc
     8101 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8161 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     8221 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8281 caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc
     8341 tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta catctacgta
     8401 ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acaccaatgg gcgtggatag
     8461 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     8521 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa
     8581 atgggcggta ggcg
//
