LOCUS       Exported                6691 bp ds-DNA     circular SYN 25-SEP-2023
DEFINITION  For mammalian expression of human ATF6 with a 3x FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    p3xFLAG-ATF6
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 Sep 25, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6691
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        556..586
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..667
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        648..666
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(705..1034)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      719..854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(773..792)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1019..1039
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1063..1685
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     1467..1486
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     CDS             complement(1705..3717)
                     /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(3721..3786)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(3816..4019)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3845..3865)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4020..4399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(4560..4577)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4560..4576)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4569..4591)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4717..5172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4854..4875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5066..5085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5199..5303
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5304..6164
                     /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(5522..5541)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6335..232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttaa tacgactcac
      661 tatagggaat taactttctg gtttttcagt tcctcgagga gctttttgca aaagcctagg
      721 cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc
      781 tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga
      841 gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc
      901 atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac
      961 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     1021 cctaactgac acacattcca cagccaagct ggccgcgtac ccaattcaac aggcatctac
     1081 tgagtggacc caacgcatga gaggacagtg ccaagcaagc aactcaaatg tcccaccggt
     1141 tgggcatggc caggtagcct atgctgtgtc tggacgtcct cctgctggta tagttatttt
     1201 aaaatcagaa ggacagggaa gggagcagtg gttcacgcct gtaatcccag caatttggga
     1261 ggccaaggtg ggtagatcac ctgagattag gagttggaga ccagcctggc caatatggtg
     1321 aaaccccgtc tctaccaaaa aaacaaaaat tagctgagcc tggtcatgca tgcctggaat
     1381 cccaacaact cgggaggctg aggcaggaga atcgcttgaa cccaggaggc ggagattgca
     1441 gtgagccaag attgtgccac tgcactccag cttggttccc aatagacccc gcaggcccta
     1501 caggttgtct tcccaacttg ccccttgctc cataccaccc ccctccaccc cataatatta
     1561 tagaaggaca cctagtcaga caaaatgatg caacttaatt ttattaggac aaggctggtg
     1621 ggcactggag tggcaacttc cagggccagg agaggcactg gggaggggtc acagggatgc
     1681 cacccgggat cctctagagt cgacctattg taatgactca gggatggtgc tgacaacgtg
     1741 ggttgcctct gtggctgcgg gaggggagcc aaagaaggtg ttggtttgat tcctctgctg
     1801 atctcggagg taaggaggaa ccgacgaact tttgatatgg aggatcctgg tgtccatcac
     1861 ctgacagtca atctgcatca tcacttcgta gtcctgccca ttgatcacat tctcatttat
     1921 gtttattgct ggtaacacaa ttgacatttt tggtcttgtg gtcttgttat gggtggtagc
     1981 tggtaacagc aggtgatccc ttcgaaatga cacaacataa aatgtgtctc cccttctgcg
     2041 gatggcttca aaaaagtctt gataacttct gggtgaagca taatacactt gtagctcact
     2101 ccctgagttc ctgctgatac tactagtggt ttctgtgtat tgaacagcca tcagctgaga
     2161 atttgagccc tgttccagag caccctgaag aatacgggtt ttctgttgat tatttgtcat
     2221 tcttcttgac ttggtccttt ctacttcatg tctatgaacc catcctcgaa gttcatgatt
     2281 taacctgaga gactctgttg tattaattag gggctgacaa ggaggtgggg gaatgtaaag
     2341 caatggttct tcagttagca ccatcagggc tttgtcattt gaaacagaat gatcatatct
     2401 gtagctgttt ttctggataa taccatctga tgtgtcctgt gcctctttag cagaaaatcc
     2461 tagaaggtgc ctcctttgat ttgcagggct cacactagga ttcattctcc tggaatcctg
     2521 ttccaacatg ctcataggtc catagttcag tattataaat gccaatacta tcatcacaca
     2581 gacaactctt cgctttggac tagggacttt aagcctctgg ttctctgaca caacttcatc
     2641 cagctgccgc ttcagtgttc cattttcttt cttcagttgc tcgttttctg agagggcagc
     2701 ctttaatctc gcctctaacc ctagcatata ttctttcttc ttcttgcgag actgacaagc
     2761 ggattctcga ttttttatta tacgttgctg tctccttagc acagcaatat ctgaaccgac
     2821 atttctcatg gtactttgta ggacaggttt agtcacggaa agttttccat tcactgggct
     2881 attcgctgaa ggggctggta ccacattcac cacgtgatta gggagctgtg tgactccccc
     2941 agcaacagca aggactggct gagcagaggg cagaactcca ggtgcttgaa gttgtaccac
     3001 agtaggctga gacagcaaaa ccgtctggcc tttagtgggt gcaggttgta aactgataat
     3061 tggttgctgc tttgccaatg gcataagcgt tggtactgtc tgaataatga tggtttttgc
     3121 tggaacactg gagtttgttt gagtcttggg tgctgctgga agcaataaag gcttgggctg
     3181 aattgaaggt tttgaattca cctgaatttt tttctttgga gtcagtccat tttcagtctt
     3241 gttcctagaa ccagtgacag gcttatcttc cttcagtggc tccggtgaag agagactatt
     3301 agagttttca ccatataagg aaaggggaga catctgagaa ctagaagaca aatccaactc
     3361 ctcaggaaca tgctgagttg aagaataaga gtccactgac cgaggagatg agactgaata
     3421 acttgaggag gctggagaaa gtggctgggg ttctgcctta atatctttaa ctgtacagat
     3481 ttggttgttg atgtcccaaa tgtctgactc ccaaggtaac aaatccaaat caaaatcaag
     3541 attatcaaaa ttgttttcat acgtctcatt tgctgcttcc aattgcagct catcagtgtc
     3601 tgtgaaataa ccgagttcag caaagagagc agaatcccaa tcttcatcca gcctgtgaaa
     3661 gagtcccggg ctaaaaggtg actccatggt gccggcaacc ccagccggct cccccataag
     3721 cttgtcatcg tcatccttgt aatcgatgtc atgatcttta taatcaccgt catggtcttt
     3781 gtagtccatg gttaattctg acggttcact aaacgagctc tgcttatata gacctcccac
     3841 cgtacacgcc taccgcccat ttgcgtcaac ggggcggggt tattacgaca ttttggaaag
     3901 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     3961 aatccccgtg agtcaaaccg ctatccacgc ccattggtgt actgccaaaa ccgcatcacc
     4021 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccgtaaggtc
     4081 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgga
     4141 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     4201 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     4261 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     4321 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     4381 ctagtcaata atcaatgtca acatggcggt catattggac atgagccaat ataaatgtac
     4441 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata
     4501 taaccaatga ctaatatggc taattgccaa tattgattca atgtatagat cagcttggca
     4561 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4621 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4681 ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt aatattttgt
     4741 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4801 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4861 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4921 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4981 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5041 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5101 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5161 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5221 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5281 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5341 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5401 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5461 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5521 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5581 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5641 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5701 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5761 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5821 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5881 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5941 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6001 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6061 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6121 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6181 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6241 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6301 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6361 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6421 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6481 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6541 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6601 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6661 gttcgtgcac acagcccagc ttggagcgaa c
//
