LOCUS       sequence-458979-        4445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient overexpression of human BEX3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4445)
  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 4445)
  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 BEX3
FEATURES             Location/Qualifiers
     source          1..4445
                     /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    497..631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(521..540)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     575..594
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(744..764)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(744..762)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(783..799)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(783..799)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(796..818)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    807..823
                     /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(831..837,838..855,856..861))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    876..897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1014..1031)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1185..1773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1265..1284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1944..2735,2736..2804))
                     /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     2567..2586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2805..2909)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2935..3390)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3072..3093)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3284..3303
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3461..4445
                     /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 gctatggcaa atattcacca ggaaaacgaa gagatggagc agcctatgca gaatggagag
      181 gaagaccgcc ctttgggagg aggtgaaggc caccagcctg caggaaatcg acggggacag
      241 gctcgccgac ttgcccctaa ttttcgatgg gccataccca ataggcagat caatgatggg
      301 atgggtggag atggagatga tatggaaata ttcatggagg agatgagaga aatcagaaga
      361 aaacttaggg agctgcagtt gaggaattgt ctgcgtatcc ttatggggga gctctctaat
      421 caccatgacc atcatgatga attttgcctt atgccttgac tcgagcctct agaactatag
      481 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
      541 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      601 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      661 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg
      721 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
      781 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      841 gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
      901 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
      961 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1021 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1081 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     1561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     1621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     1681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     1741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     1801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     1861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     2941 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3001 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3061 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3121 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3181 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3241 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3301 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3361 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     3421 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc aattcaatat
     3481 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat
     3541 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
     3601 ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc aactggggag
     3661 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
     3721 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
     3781 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
     3841 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
     3901 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
     3961 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
     4021 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
     4081 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
     4141 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
     4201 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaagtacg
     4261 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
     4321 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
     4381 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
     4441 ggaac
//
