LOCUS       sequence-335159-        6840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses of ATF6-alpha(1-373) in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  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 6840)
  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: pET41SIII-ATF6(1-373)
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(150..173)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(1311..1334)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(1353..2006)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(1373..1395)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     RBS             2014..2036
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2051..2075
                     /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)."
     primer_bind     complement(2075..2094)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2076..2094)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2182..2201)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2407..2484
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2485..3567
                     /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     complement(2504..2523)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4139..4330
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4324..4346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4432..4571
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4487..4506
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4586..4603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4757..5345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4837..4856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5456..5475)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5467..6282
                     /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             5467..6282
                     /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     complement(5540..5559)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6374..6829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6511..6532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6723..6742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgacg
      181 gatccactag ggactttaag cctctggttc tctgacacaa cttcatccag ctgccgcttc
      241 agtgttccat tttctttctt cagttgctcg ttttctgaga gggcagcctt taatctcgcc
      301 tctaacccta gcatatattc tttcttcttc ttgcgagact gacaagcgga ttctcgattt
      361 tttatcatac gttgctgtct ccttagcaca gcaatatctg aaccgacatt tctcatggta
      421 ctttgtagga caggtttagt cacggaaagt tttccattca ctgggctatt cgctgaaggg
      481 gctggtacca cattcaccac gtgattaggg agctgtgtga ctcccccagc aacagcaagg
      541 actggctgag cagagggcag aactccaggt gcttgaagtt gtaccacagt aggctgagac
      601 agcaaaaccg tctggccttt agtgggtgca ggttgtaaac tgataattgg ttgctgcttt
      661 gccaatggca taagcgttgg tactgtctga ataatgatgg tttttgctgg aacactggag
      721 tttgtttgag tcttgggtgc tgctggaagc aataaaggct tgggctgaat tgaaggtttt
      781 gaattcacct gaattttttt ctttggagtc agtccatttt cagtcttgtt cctaggacca
      841 gtgacaggct tatcttcctt cagtggctcc gctgaagaga gactattaga gttttcacca
      901 tataaggaaa ggggagacat ctgagaacta gaagacaaat ccaactcctc aggaacatgc
      961 tgagttgaag aataagagtc cactgaccga ggagatgaga ctgaataact tgaggaggct
     1021 ggagaaagtg gctggggttc tgccttaata tctttaactg tacagatttg gttgttgatg
     1081 tcccaaatgt ctgactccca aggcatcaaa tccaaatcaa aatcaagatt atcaaaattg
     1141 ttttcatacg tctcatttgc tgcttccaat tgcagctcat cagtgtctgt gaaataacca
     1201 agttcagcaa agagagcaga atcccaatct tcatccagcc tgtgaaagag tcccgggcta
     1261 aaaggtgact ccatggtgcc ggcaacccca gccggctccc ccatctcgag gggcccctgg
     1321 aacagaactt ccagactagt tgaaccatcc gattttggag gatggtcgcc accaccaaac
     1381 gtggcttgcc agccctgcaa aggccatgct atatacttgc tggatttcaa gtacttatca
     1441 atttgtggga tagcttcaat acgtttttta aaacaaacta attttgggaa cgcatccagg
     1501 cacattgggt ccatgtataa aacaacatca agagcgtcat acaacatgaa gtcaggatgg
     1561 gttacatgat caccatttaa atatgtttta tgacataaac gatcttcgaa cattttcagc
     1621 atttcaggta gcttgctaag aaaatcaact ttgagagttt caaagtcttt actatatgca
     1681 attctcgaaa caccgtatct aatatccaaa accgctcctt caagcattga aatctctgca
     1741 cgctcttttg gacaaccacc caacatgttg tgcttgtcag ctatataacg tatgatggcc
     1801 atagactgtg ttaatttaac atcaccatca atataataag gaagattggg aaactccaaa
     1861 cccaattcaa actttttgtt tcgccattta tcaccttcat cgcgctcata caaatgctct
     1921 tcatattttt cttcaagata ttccaaaaga agtcgagtgg gttgcacaag gcccttaatt
     1981 ttccaataac ctagtatagg ggacatatgt atatctcctt cttaaagtta aacaaaatta
     2041 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     2101 cgggatcgag atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
     2161 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     2221 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     2281 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     2341 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
     2401 atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     2461 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     2521 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     2581 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     2641 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     2701 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     2761 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     2821 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     2881 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     2941 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     3001 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     3061 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     3121 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     3181 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     3241 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga
     3301 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     3361 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     3421 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     3481 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3541 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     3601 ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     3661 gtgggcgcgg ggcatgacta gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg
     3721 gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga
     3781 ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt
     3841 ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat
     3901 cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca
     3961 ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc
     4021 acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc
     4081 tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt
     4141 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
     4201 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
     4261 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
     4321 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4381 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4441 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     4501 attgtactga gagtgcacca ttgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4561 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4621 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4681 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4741 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4801 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4861 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4921 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4981 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5041 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5101 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5161 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5221 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5281 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5341 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5401 gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     5461 ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac
     5521 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     5581 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     5641 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     5701 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     5761 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     5821 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     5881 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     5941 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     6001 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     6061 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     6121 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     6181 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     6241 aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg
     6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6361 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     6421 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     6481 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     6541 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     6601 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     6661 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     6721 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6781 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
