LOCUS       sequence-335508-        7317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expresses of Delta-N-ELOA containing Elongin complex in bacteria.
            The resulting plasmid was used to generate a single expression 
            construct encoding all 3Elongin subunits, with a 6-His tag on ELOB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7317)
  AUTHORS   He Y, Sato S, Tomomori-Sato C, Chen S, Goode ZH, Conaway JW, Conaway
            RC
  TITLE     Elongin functions as a loading factor for Mediator at 
            ATF6alpha-regulated ER stress response genes.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Sep 28;118(39):e2108751118. doi: 
            10.1073/pnas.2108751118.
  PUBMED    34544872
REFERENCE   2  (bases 1 to 7317)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-09-28"; volume: "118(39):e2108751118. 
            doi"; pages: " 10.1073/pnas.2108751118"
COMMENT     Sequence Label: pQ-Elo-HB-C-A(400-772)
FEATURES             Location/Qualifiers
     source          1..7317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(36..50,51..56,57..73,74..80)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    56..72
                     /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)."
     protein_bind    88..104
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             147..167
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(571..595)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      617..711
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(740..754,755..760,761..777,778..784)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    760..776
                     /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)."
     protein_bind    792..808
                     /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)."
     RBS             827..838
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(1224..1248)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      1270..1364
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        join(1393..1407,1408..1413,1414..1430,1431..1437)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1413..1429
                     /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)."
     protein_bind    1445..1461
                     /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)."
     RBS             1480..1491
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(2663..2687)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      2709..2803
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2912..3571
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2978..2997)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3636..3722
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3815..4897)
                     /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     4859..4878
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4898..4975)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5059..5081)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5167..5307
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5222..5241
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5322..5339)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5493..6081)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5573..5592)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6252..7043,7044..7112))
                     /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     6875..6894
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7113..7217)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7285..7303
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagctta attaacaaca ccatttgtcg agaaatcata aaaaatttat ttgctttgtg
       61 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      121 ttaaagagga gaaattaact atgaaacatc accatcacca tcaccatggt gctatggacg
      181 tgttcctcat gatccggcgc cacaagacca ccatcttcac ggacgccaag gagtccagca
      241 cggtgttcga actgaagcgc atcgtcgagg gcatcctcaa gcggcctcct gacgagcagc
      301 ggctgtacaa ggatgaccaa ctcttggatg atggcaagac actgggcgag tgtggcttca
      361 ccagtcaaac agcacggcca caggccccag ccacagtggg gctggccttc cgggcagatg
      421 acacctttga ggccctgtgc atcgagccgt tttccagccc gccagagctg cccgatgtga
      481 tgaagcccca ggactcggga agcagtgcca atgaacaagc cgtgcagtga ggatccagtc
      541 ttcgcatgca tgaagacgcg gccgcctagg acccagcttt cttgtacaaa gtggtaagct
      601 taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
      661 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagtaac
      721 aacaccattt gtcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat
      781 aatagattca attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt
      841 aactatggat ggagaggaga aaacctatgg tggctgtgaa ggacctgatg ccatgtatgt
      901 caaattgata tcatctgatg gccatgaatt tattgtaaaa agagaacatg cattaacatc
      961 aggcacgata aaagccatgt tgagtggccc aggtcagttt gctgagaacg aaaccaatga
     1021 ggtcaatttt agagagatac cttcacatgt gctatcgaaa gtatgcatgt attttacgta
     1081 caaggttcgc tacactaaca gctccaccga gattcctgaa ttcccaattg cacctgaaat
     1141 tgcactggaa ctgctgatgg ctgcgaactt cttagattgt taaggatcca gtcttcgcat
     1201 gcatgaagac gcggccgcct aggacccagc tttcttgtac aaagtggtaa gcttaattag
     1261 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1321 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagt aacaacacca
     1381 tttgtcgaga aatcataaaa aatttatttg ctttgtgagc ggataacaat tataatagat
     1441 tcaattgtga gcggataaca atttcacaca gaattcatta aagaggagaa attaaccatg
     1501 ttcgagcagc caaccatgtc ttttgaatcc tacctcagct atgaccagcc ccggaagaaa
     1561 aagaaaaaga ttgtgaaaac ttcagccacg gcacttggag ataaaggact taaaaaaaat
     1621 gactctaaaa gcactggtaa aaacttggac tcagttcaga aattacccaa ggtgaacaaa
     1681 accaagtcag agaagccggc tggagctgat ttagccaagc tgagaaaggt gcctgatgtg
     1741 ttgccagtgt tgccagacct cccgttaccc gcgatacagg ccaattaccg tccactgcct
     1801 tccctcgagc tgatatcctc cttccagcca aagcgaaaag cgttctcttc accccaggaa
     1861 gaagaagaag ctggatttac tgggcgcaga atgaattcca agatgcaggt gtattctggt
     1921 tccaagtgtg cctatctccc taaaatgatg accttgcacc agcaatgcat ccgagtactt
     1981 aaaaacaaca tcgattcaat ctttgaagtg ggaggagtcc catactctgt tcttgaaccc
     2041 gttttggaga ggtgtacacc tgatcagctg tatcgcatag aggaatacaa tcatgtatta
     2101 attgaagaaa cagatcaatt atggaaagtt cattgtcacc gagactttaa ggaagaaaga
     2161 cccgaagagt atgagtcgtg gcgagagatg tacctgcggc ttcaggacgc ccgagagcag
     2221 cggctacgag tactaacaaa gaatatccag ttcgcacatg ccaataagcc caaaggccga
     2281 caagcaaaga tggcctttgt caactctgtg gccaagccac ctcgtgacgt ccggaggagg
     2341 caggaaaagt ttggaacggg aggagcagct gtccctgaga aaatcaagat caagccagcc
     2401 ccgtacccca tgggaagcag ccatgcttcc gccagtagca tcagctttaa ccccagccct
     2461 gaggagccgg cctatgatgg cccaagcacc agcagtgccc acttggcacc agtggtcagc
     2521 agcactgttt cctatgatcc taggaaaccc actgtgaaga aaattgcccc aatgatggcc
     2581 aagacaatta aagctttcaa gaacagattc tcccgacgat aaggatccag tcttcgcatg
     2641 catgaagacg cggccgccta ggacccagct ttcttgtaca aagtggtaag cttaattagc
     2701 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     2761 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagta acaacaccat
     2821 ttaaatggag tggttacaaa tggagtggtt acaaatggag tggttaatta aggctagctt
     2881 ggcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     2941 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     3001 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     3061 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     3121 tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     3181 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     3241 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     3301 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     3361 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     3421 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     3481 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     3541 gtactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     3601 aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga
     3661 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     3721 atccgccctc tagattacgt gcagtcgatg ataagctgtc aaacatgaga attgtgccta
     3781 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     3961 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4021 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4081 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4141 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     4201 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     4261 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     4321 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     4381 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     4441 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     4501 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     4561 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     4621 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     4681 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     4741 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     4801 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     4861 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     4921 cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg gtgtcggaat
     4981 ttcgggcagc gttgggtcct ggccacgggt gcgcatgatc tagagctgcc tcgcgcgttt
     5041 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     5101 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     5161 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     5221 gcggcatcag agcagattgt actgagagtg caccacatgc ggtgtgaaat accgcacaga
     5281 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     5341 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5401 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5461 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5521 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5581 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5641 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5701 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5761 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5821 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5881 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5941 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6001 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6061 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6121 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6181 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6241 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6301 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6361 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6421 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6481 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6541 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6601 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6661 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6721 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6781 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6841 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6901 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6961 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7021 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7081 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7141 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7201 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7261 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcac
//
