LOCUS       sequence-458978-        4479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient overexpression of human BEX1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4479)
  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 4479)
  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-BEX1
FEATURES             Location/Qualifiers
     source          1..4479
                     /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    531..665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(555..574)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     609..628
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(778..798)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(778..796)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(817..833)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(817..833)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(830..852)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    841..857
                     /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(865..871,872..889,890..895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    910..931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1048..1065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1219..1807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1299..1318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1978..2769,2770..2838))
                     /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     2601..2620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2839..2943)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2969..3424)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3127)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3318..3337
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3495..4479
                     /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 ccaaagagaa acgagcagta aacagtctca gcatggaaaa tgccaaccaa
      181 gaaaatgaag aaaaggagca agttgctaat aaaggggagc ccttggccct ccctttggat
      241 gctggtgaat actgtgtgcc tagaggaaat cgtaggcggt tccgcgttag gcagcccatc
      301 ctgcagtata gatgggatat gatgcatagg cttggagaac cacaggcaag gatgagagaa
      361 gagaatatgg aaaggattgg ggaggaggtg agacagctga tggaaaagct gagggaaaag
      421 cagttgagtc atagtctgcg ggcagtcagc actgaccccc ctcaccatga ccatcatgat
      481 gagttttgcc ttatgccctg atctagaact atagtgagtc gtattacgta gatccagaca
      541 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      601 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      661 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      721 ttttttaatt cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct
      781 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
      841 ttgttatccg ctcacaattc cacacaacat acgagccggg agcataaagt gtaaagcctg
      901 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
      961 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1021 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1081 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1141 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1201 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1261 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1321 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1381 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1441 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1501 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1561 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1621 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1681 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1741 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1801 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1861 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1921 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1981 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2041 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2101 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2161 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2221 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2281 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2341 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     2401 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2461 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2521 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2581 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2641 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2701 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2761 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2821 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2881 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2941 gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc
     3001 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     3061 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     3121 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     3181 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     3241 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     3301 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     3361 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3421 gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     3481 tcgctattac gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca
     3541 atatggtgga tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca
     3601 atatggtgga tctggacccc agccaattca atatggcgga cttggcacca tgccaattca
     3661 atatggcgga cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt
     3721 tgaggagggg tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa
     3781 tggcggccat attggctata tgccaggatc aatatatagg caatatccaa tatggcccta
     3841 tgccaatatg gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt
     3901 ccatatatgg gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc
     3961 atatatgggg cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta
     4021 tatatggtct ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc
     4081 ccattgacgt caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg
     4141 accacccacc attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc
     4201 ccctattgac gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata
     4261 gtaacttggc aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat
     4321 tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa
     4381 tggggagggg caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc
     4441 taatgggagg tctatataag caatgctcgt ttagggaac
//
