LOCUS       sequence-458859-        4464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient overexpression of human BEX4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4464)
  AUTHORS   Manford AG, Mena EL, Shih KY, Gee CL, McMinimy R, Martinez-Gonzalez 
            B, Sherriff R, Lew B, Zoltek M, Rodriguez-Perez F, Woldesenbet M, 
            Kuriyan J, Rape M
  TITLE     Structural basis and regulation of the reductive stress response.
  JOURNAL   Cell. 2021 Oct 14;184(21):5375-5390.e16. doi: 
            10.1016/j.cell.2021.09.002. Epub 2021 Sep 24.
  PUBMED    34562363
REFERENCE   2  (bases 1 to 4464)
  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.cell.2021.09.002"; journalName: "Cell"; date: 
            "2021-10-14- 14"; volume: "184"; issue: "21"; pages: "5375-5390.e16"
COMMENT     Sequence Label: pCS2+ HA-BEX4
FEATURES             Location/Qualifiers
     source          1..4464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             97..123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    516..650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(540..559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     594..613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(763..783)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(763..781)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(802..818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(815..837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    826..842
                     /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(850..856,857..874,875..880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    895..916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1033..1050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1204..1792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1284..1303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1963..2754,2755..2823))
                     /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     2586..2605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2824..2928)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2954..3409)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3091..3112)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3303..3322
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3480..4464
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccacc atgggctacc catacgatgt tccagattac
      121 gctatggagt ccaaagagga actagcggca aacaatctca acggggaaaa tgcccaacaa
      181 gaaaacgaag gaggggagca ggcccccacg cagaatgaag aagaatcccg ccatttggga
      241 gggggtgaag gccagaagcc tggaggaaat atcaggcggg ggcgagttag gcgacttgtc
      301 cctaattttc gatgggccat acctaatagg catattgagc acaatgaagc gagagatgat
      361 gtagaaaggt ttgtagggca gatgatggaa atcaagagaa agactaggga acagcagatg
      421 aggcactata tgcgcttcca aactcctgaa cctgacaacc attatgactt ttgcctcata
      481 ccttgatcta gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg
      541 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      601 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      661 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg
      721 ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat
      781 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      841 aattccacac aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt
      901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2941 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3001 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3061 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3121 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3181 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3241 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3301 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3361 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     3421 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3481 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
     3541 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
     3601 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
     3661 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
     3721 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
     3781 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
     3841 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
     3901 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
     3961 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
     4021 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
     4081 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
     4141 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
     4201 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
     4261 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
     4321 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
     4381 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     4441 ataagcaatg ctcgtttagg gaac
//
