LOCUS       sequence-458871-        4405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression vector containing mouse ElonginB and ElonginC for 
            E.coli protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4405)
  AUTHORS   Manford AG, Rodriguez-Perez F, Shih KY, Shi Z, Berdan CA, Choe M, 
            Titov DV, Nomura DK, Rape M
  TITLE     A Cellular Mechanism to Detect and Alleviate Reductive Stress.
  JOURNAL   Cell. 2020 Oct 1;183(1):46-61.e21. doi: 10.1016/j.cell.2020.08.034. 
            Epub 2020 Sep 16.
  PUBMED    32941802
REFERENCE   2  (bases 1 to 4405)
  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.2020.08.034"; journalName: "Cell"; date: "2020-10-1-
            1"; volume: "183"; issue: "1"; pages: "46-61.e21"
COMMENT     Sequence Label: pRSFDuet-1 ElonginB, ElonginC
FEATURES             Location/Qualifiers
     source          1..4405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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        505..523
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    524..548
                     /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)."
     CDS             942..986
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1024..1042)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1038..1085
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1315..2130)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1315..2130)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2038..2057
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2131..2222)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2238..2987)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(2248..2265)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2488..2507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3183..4265)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4227..4246
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4266..4343)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4389..4405,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggacgtgt ttctcatgat ccggcgccac aagaccacca tctttacgga
      121 cgccaaggag tcgagcaccg tgttcgaact gaagcgcatc gtcgagggca tcctcaagcg
      181 gccgccagag gagcagcggc tttacaagga tgaccagctc cttgatgatg gcaaaactct
      241 gggcgagtgt ggcttcacta gccagacagc acggccacag gccccagcca cagtgggcct
      301 ggccttccga gcagatgaca ccttcgaagc tctccgcatc gagccctttt ccagccctcc
      361 agagcttcca gatgtgatga agccacagga ttctggaggc agtgccaatg aacaagctgt
      421 gcagtgagtc gacaagcttg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat
      481 tgtacacggc cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata
      541 acaattcccc atcttagtat attagttaag tataagaagg agatatacat atggatggag
      601 aggagaagac ctatggtggc tgtgaaggcc ctgatgctat gtatgtgaaa ttaatatctt
      661 ctgatggcca tgaatttatt gtaaaaagag aacatgcact aacatcagga acaataaagg
      721 ctatgttgag tggtccaggt cagtttgctg agaatgaaac caacgaggtc aattttagag
      781 agatcccctc acatgtgcta tcaaaagtgt gcatgtattt tacctacaag gtccgctata
      841 ctaacagctc cactgaaatt cctgaattcc caattgcacc tgaaattgca ctggaactgc
      901 tgatggctgc aaacttccta gattgttaac tcgagtctgg taaagaaacc gctgctgcga
      961 aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg
     1021 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1081 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     1141 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     1201 caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct
     1261 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     1321 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     1381 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     1441 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     1501 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     1561 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     1621 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     1681 agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     1741 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     1801 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     1861 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     1921 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     1981 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     2041 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     2101 gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat
     2161 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2221 aataggcatg cagcgctctt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     2281 actgcggcga gcggtgtcag ctcactcaaa agcggtaata cggttatcca cagaatcagg
     2341 ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag ccaacgccgc
     2401 aggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagcc
     2461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2581 cgggaagcgt ggcgctttct catagctcac gctgttggta tctcagttcg gtgtaggtcg
     2641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2761 ccattggtaa ctgatttaga ggactttgtc ttgaagttat gcacctgtta aggctaaact
     2821 gaaagaacag attttggtga gtgcggtcct ccaacccact taccttggtt caaagagttg
     2881 gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg tggtttttct ttatttatga
     2941 gatgatgaat caatcggtct atcaagtcaa cgaacagcta ttccgttact ctagatttca
     3001 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     3061 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     3121 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     3181 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3241 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     3301 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     3361 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     3421 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     3481 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     3541 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     3601 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     3661 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     3721 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     3781 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     3841 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     3901 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     3961 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     4021 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     4081 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     4141 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     4201 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     4261 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     4321 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     4381 taggaaatta atacgactca ctata
//
