LOCUS       sequence-442102-        6691 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian expression of human ATF6 with a 3x FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6691)
  AUTHORS   Chen X, Shen J, Prywes R
  TITLE     The luminal domain of ATF6 senses endoplasmic reticulum (ER) stress 
            and causes translocation of ATF6 from the ER to the Golgi.
  JOURNAL   J Biol Chem. 2002 Apr 12. 277(15):13045-52.
  PUBMED    11821395
REFERENCE   2  (bases 1 to 6691)
  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: "J Biol 
            Chem. 2002 Apr 12. 277(15):13045-52."
COMMENT     Sequence Label: p3xFLAG-ATF6
FEATURES             Location/Qualifiers
     source          1..6691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     241..261
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    285..907
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     689..708
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(927..2939)
                     /codon_start=1
                     /product="human activating transcription factor 6, an 
                     endoplasmic reticulum (ER) protein involved in the unfolded
                     protein response"
                     /label=ATF6
                     /note="Accumulation of misfolded proteins in the ER leads 
                     to proteolytic cleavage that releases the cytosolic domain 
                     of ATF6, which enters the nucleus and acts as a 
                     transcription factor."
                     /translation="MGEPAGVAGTMESPFSPGLFHRLDEDWDSALFAELGYFTDTDELQ
                     LEAANETYENNFDNLDFDLDLLPWESDIWDINNQICTVKDIKAEPQPLSPASSSYSVSS
                     PRSVDSYSSTQHVPEELDLSSSSQMSPLSLYGENSNSLSSPEPLKEDKPVTGSRNKTEN
                     GLTPKKKIQVNSKPSIQPKPLLLPAAPKTQTNSSVPAKTIIIQTVPTLMPLAKQQPIIS
                     LQPAPTKGQTVLLSQPTVVQLQAPGVLPSAQPVLAVAGGVTQLPNHVVNVVPAPSANSP
                     VNGKLSVTKPVLQSTMRNVGSDIAVLRRQQRIIKNRESACQSRKKKKEYMLGLEARLKA
                     ALSENEQLKKENGTLKRQLDEVVSENQRLKVPSPKRRVVCVMIVLAFIILNYGPMSMLE
                     QDSRRMNPSVSPANQRRHLLGFSAKEAQDTSDGIIQKNSYRYDHSVSNDKALMVLTEEP
                     LLYIPPPPCQPLINTTESLRLNHELRGWVHRHEVERTKSRRMTNNQQKTRILQGALEQG
                     SNSQLMAVQYTETTSSISRNSGSELQVYYASPRSYQDFFEAIRRRGDTFYVVSFRRDHL
                     LLPATTHNKTTRPKMSIVLPAININENVINGQDYEVMMQIDCQVMDTRILHIKSSSVPP
                     YLRDQQRNQTNTFFGSPPAATEATHVVSTIPESLQ"
     CDS             complement(2943..3008)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3038..3241)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3067..3087)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3242..3621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3782..3799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3782..3798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3791..3813)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3939..4394)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4076..4097)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4288..4307
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4421..4525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4526..4594,4595..5386)
                     /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     complement(4744..4763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5557..6145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6046..6065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6299..6316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6433..6454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6469..6474,6475..6492,6493..6499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6507..6523
                     /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     6512..6534
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6531..6547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6531..6547
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6561..6580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6561..6579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(6618..6691,1..256))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6632..6691,1..76)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
       61 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
      121 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      181 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      241 accctaactg acacacattc cacagccaag ctggccgcgt acccaattca acaggcatct
      301 actgagtgga cccaacgcat gagaggacag tgccaagcaa gcaactcaaa tgtcccaccg
      361 gttgggcatg gccaggtagc ctatgctgtg tctggacgtc ctcctgctgg tatagttatt
      421 ttaaaatcag aaggacaggg aagggagcag tggttcacgc ctgtaatccc agcaatttgg
      481 gaggccaagg tgggtagatc acctgagatt aggagttgga gaccagcctg gccaatatgg
      541 tgaaaccccg tctctaccaa aaaaacaaaa attagctgag cctggtcatg catgcctgga
      601 atcccaacaa ctcgggaggc tgaggcagga gaatcgcttg aacccaggag gcggagattg
      661 cagtgagcca agattgtgcc actgcactcc agcttggttc ccaatagacc ccgcaggccc
      721 tacaggttgt cttcccaact tgccccttgc tccataccac ccccctccac cccataatat
      781 tatagaagga cacctagtca gacaaaatga tgcaacttaa ttttattagg acaaggctgg
      841 tgggcactgg agtggcaact tccagggcca ggagaggcac tggggagggg tcacagggat
      901 gccacccggg atcctctaga gtcgacctat tgtaatgact cagggatggt gctgacaacg
      961 tgggttgcct ctgtggctgc gggaggggag ccaaagaagg tgttggtttg attcctctgc
     1021 tgatctcgga ggtaaggagg aaccgacgaa cttttgatat ggaggatcct ggtgtccatc
     1081 acctgacagt caatctgcat catcacttcg tagtcctgcc cattgatcac attctcattt
     1141 atgtttattg ctggtaacac aattgacatt tttggtcttg tggtcttgtt atgggtggta
     1201 gctggtaaca gcaggtgatc ccttcgaaat gacacaacat aaaatgtgtc tccccttctg
     1261 cggatggctt caaaaaagtc ttgataactt ctgggtgaag cataatacac ttgtagctca
     1321 ctccctgagt tcctgctgat actactagtg gtttctgtgt attgaacagc catcagctga
     1381 gaatttgagc cctgttccag agcaccctga agaatacggg ttttctgttg attatttgtc
     1441 attcttcttg acttggtcct ttctacttca tgtctatgaa cccatcctcg aagttcatga
     1501 tttaacctga gagactctgt tgtattaatt aggggctgac aaggaggtgg gggaatgtaa
     1561 agcaatggtt cttcagttag caccatcagg gctttgtcat ttgaaacaga atgatcatat
     1621 ctgtagctgt ttttctggat aataccatct gatgtgtcct gtgcctcttt agcagaaaat
     1681 cctagaaggt gcctcctttg atttgcaggg ctcacactag gattcattct cctggaatcc
     1741 tgttccaaca tgctcatagg tccatagttc agtattataa atgccaatac tatcatcaca
     1801 cagacaactc ttcgctttgg actagggact ttaagcctct ggttctctga cacaacttca
     1861 tccagctgcc gcttcagtgt tccattttct ttcttcagtt gctcgttttc tgagagggca
     1921 gcctttaatc tcgcctctaa ccctagcata tattctttct tcttcttgcg agactgacaa
     1981 gcggattctc gattttttat tatacgttgc tgtctcctta gcacagcaat atctgaaccg
     2041 acatttctca tggtactttg taggacaggt ttagtcacgg aaagttttcc attcactggg
     2101 ctattcgctg aaggggctgg taccacattc accacgtgat tagggagctg tgtgactccc
     2161 ccagcaacag caaggactgg ctgagcagag ggcagaactc caggtgcttg aagttgtacc
     2221 acagtaggct gagacagcaa aaccgtctgg cctttagtgg gtgcaggttg taaactgata
     2281 attggttgct gctttgccaa tggcataagc gttggtactg tctgaataat gatggttttt
     2341 gctggaacac tggagtttgt ttgagtcttg ggtgctgctg gaagcaataa aggcttgggc
     2401 tgaattgaag gttttgaatt cacctgaatt tttttctttg gagtcagtcc attttcagtc
     2461 ttgttcctag aaccagtgac aggcttatct tccttcagtg gctccggtga agagagacta
     2521 ttagagtttt caccatataa ggaaagggga gacatctgag aactagaaga caaatccaac
     2581 tcctcaggaa catgctgagt tgaagaataa gagtccactg accgaggaga tgagactgaa
     2641 taacttgagg aggctggaga aagtggctgg ggttctgcct taatatcttt aactgtacag
     2701 atttggttgt tgatgtccca aatgtctgac tcccaaggta acaaatccaa atcaaaatca
     2761 agattatcaa aattgttttc atacgtctca tttgctgctt ccaattgcag ctcatcagtg
     2821 tctgtgaaat aaccgagttc agcaaagaga gcagaatccc aatcttcatc cagcctgtga
     2881 aagagtcccg ggctaaaagg tgactccatg gtgccggcaa ccccagccgg ctcccccata
     2941 agcttgtcat cgtcatcctt gtaatcgatg tcatgatctt tataatcacc gtcatggtct
     3001 ttgtagtcca tggttaattc tgacggttca ctaaacgagc tctgcttata tagacctccc
     3061 accgtacacg cctaccgccc atttgcgtca acggggcggg gttattacga cattttggaa
     3121 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     3181 gaaatccccg tgagtcaaac cgctatccac gcccattggt gtactgccaa aaccgcatca
     3241 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccgtaagg
     3301 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     3361 gacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     3421 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3481 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3541 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     3601 aactagtcaa taatcaatgt caacatggcg gtcatattgg acatgagcca atataaatgt
     3661 acatattatg atatagatac aacgtatgca atggccaata gccaatattg atttatgcta
     3721 tataaccaat gactaatatg gctaattgcc aatattgatt caatgtatag atcagcttgg
     3781 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3841 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3901 gcccttccca acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt
     3961 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4021 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4081 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4141 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4201 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4261 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4321 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4381 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4441 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4501 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4561 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4621 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4681 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4741 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4801 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4861 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4921 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4981 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5041 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5101 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5161 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5221 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5281 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5341 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5401 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5461 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5521 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5581 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5641 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5701 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5761 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5821 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5881 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5941 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6001 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6061 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6121 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6181 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6241 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6301 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6361 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6421 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     6481 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     6541 ctatgacatg attacgaatt taatacgact cactataggg aattaacttt ctggtttttc
     6601 agttcctcga ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     6661 tggaatagct cagaggccga ggcggcctcg g
//
