LOCUS       sequence-226826-        6729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For visualization of ATF6 in mammalian cells with a mutation in ATF6
            that blocks cleavage by protease S1P. Activated protein will remain 
            in the Golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6729)
  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 6729)
  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: pEGFP-ATF6-(S1P-)
FEATURES             Location/Qualifiers
     source          1..6729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(671..673,674..676,677..1387)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(716..737)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(977..996)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1324..1345
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1412..3424
                     /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
                     QDSRRMNPSVSPANQAAHLLGFSAKEAQDTSDGIIQKNSYRYDHSVSNDKALMVLTEEP
                     LLYIPPPPCQPLINTTESLRLNHELRGWVHRHEVERTKSRRMTNNQQKTRILQGALEQG
                     SNSQLMAVQYTETTSSISRNSGSELQVYYASPRSYQDFFEAIRRRGDTFYVVSFRRDHL
                     LLPATTHNKTTRPKMSIVLPAININENVINGQDYEVMMQIDCQVMDTRILHIKSSSVPP
                     YLRDQQRNQTNTFFGSPPAATEATHVVSTIPESLQ"
     polyA_signal    3575..3696
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3612..3631)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3666..3685
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3703..4158)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3840..3861)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4052..4071
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4185..4289
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4291..4648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4291..4482
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4314..4334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4499..4634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4683..5477
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4683..5477
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4737..4756)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5347..5366
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5665..5684)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5709..5756
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6085..6673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6574..6593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
      721 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
      781 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
      841 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
      901 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
      961 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1021 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1081 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1141 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1201 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1261 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1321 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1381 gtacaagtac tcagatctcg agctcaagct tatgggggag ccggctgggg ttgccggcac
     1441 catggagtca ccttttagcc cgggactctt tcacaggctg gatgaagatt gggattctgc
     1501 tctctttgct gaactcggtt atttcacaga cactgatgag ctgcaattgg aagcagcaaa
     1561 tgagacgtat gaaaacaatt ttgataatct tgattttgat ttggatttgt taccttggga
     1621 gtcagacatt tgggacatca acaaccaaat ctgtacagtt aaagatatta aggcagaacc
     1681 ccagccactt tctccagcct cctcaagtta ttcagtctca tctcctcggt cagtggactc
     1741 ttattcttca actcagcatg ttcctgagga gttggatttg tcttctagtt ctcagatgtc
     1801 tcccctttcc ttatatggtg aaaactctaa tagtctctct tcaccggagc cactgaagga
     1861 agataagcct gtcactggtt ctaggaacaa gactgaaaat ggactgactc caaagaaaaa
     1921 aattcaggtg aattcaaaac cttcaattca gcccaagcct ttattgcttc cagcagcacc
     1981 caagactcaa acaaactcca gtgttccagc aaaaaccatc attattcaga cagtaccaac
     2041 gcttatgcca ttggcaaagc agcaaccaat tatcagttta caacctgcac ccactaaagg
     2101 ccagacggtt ttgctgtctc agcctactgt ggtacaactt caagcacctg gagttctgcc
     2161 ctctgctcag ccagtccttg ctgttgctgg gggagtcaca cagctcccta atcacgtggt
     2221 gaatgtggta ccagcccctt cagcgaatag cccagtgaat ggaaaacttt ccgtgactaa
     2281 acctgtccta caaagtacca tgagaaatgt cggttcagat attgctgtgc taaggagaca
     2341 gcaacgtata ataaaaaatc gagaatccgc ttgtcagtct cgcaagaaga agaaagaata
     2401 tatgctaggg ttagaggcga gattaaaggc tgccctctca gaaaacgagc aactgaagaa
     2461 agaaaatgga acactgaagc ggcagctgga tgaagttgtg tcagagaacc agaggcttaa
     2521 agtccctagt ccaaagcgaa gagttgtctg tgtgatgata gtattggcat ttataatact
     2581 gaactatgga cctatgagca tgttggaaca ggattccagg agaatgaatc ctagtgtgag
     2641 ccctgcaaat caagcggcgc accttctagg attttctgct aaagaggcac aggacacatc
     2701 agatggtatt atccagaaaa acagctacag atatgatcat tctgtttcaa atgacaaagc
     2761 cctgatggtg ctaactgaag aaccattgct ttacattccc ccacctcctt gtcagcccct
     2821 aattaataca acagagtctc tcaggttaaa tcatgaactt cgaggatggg ttcatagaca
     2881 tgaagtagaa aggaccaagt caagaagaat gacaaataat caacagaaaa cccgtattct
     2941 tcagggtgct ctggaacagg gctcaaattc tcagctgatg gctgttcaat acacagaaac
     3001 cactagtagt atcagcagga actcagggag tgagctacaa gtgtattatg cttcacccag
     3061 aagttatcaa gacttttttg aagccatccg cagaagggga gacacatttt atgttgtgtc
     3121 atttcgaagg gatcacctgc tgttaccagc taccacccat aacaagacca caagaccaaa
     3181 aatgtcaatt gtgttaccag caataaacat aaatgagaat gtgatcaatg ggcaggacta
     3241 cgaagtgatg atgcagattg actgtcaggt gatggacacc aggatcctcc atatcaaaag
     3301 ttcgtcggtt cctccttacc tccgagatca gcagaggaat caaaccaaca ccttctttgg
     3361 ctcccctccc gcagccacag aggcaaccca cgttgtcagc accatccctg agtcattaca
     3421 ataggtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     3481 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3541 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3601 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3661 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3721 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3781 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3841 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3901 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3961 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4021 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4081 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4141 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4201 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4261 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4321 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4381 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4441 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4501 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4561 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4621 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4681 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4741 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4801 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4861 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4921 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4981 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5041 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5101 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5161 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5221 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5281 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5341 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5401 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5461 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5521 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5581 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5641 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5701 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5761 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5821 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5881 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5941 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6001 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6061 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6121 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6181 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6541 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6601 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6661 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6721 ttttgctca
//
