LOCUS       sequence-173332-        6729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For visualization of GFP-ATF6 in mammalian cells..
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
FEATURES             Location/Qualifiers
     source          1..6729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        69..426
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        69..260
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(92..112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      277..412
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             461..1255
                     /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             461..1255
                     /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(515..534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1125..1144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1443..1462)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1487..1534
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1863..2451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2352..2371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2626..2929
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2930..3133
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3084..3104
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3172..3181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3178..3180,3181..3183,3184..3894)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3178..3180,3181..3183,3184..3894)
                     /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(3223..3244)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3484..3503)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3831..3852
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3919..5931
                     /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"
     polyA_signal    6082..6203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6119..6138)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6173..6192
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6210..6665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6347..6368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6559..6578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6692..6729,1..67)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
       61 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      121 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      181 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      241 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      301 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      361 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      421 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      481 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      541 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      601 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
      661 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      721 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      781 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      841 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      901 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      961 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     1021 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1081 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1141 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1201 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1261 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     1321 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1381 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     1441 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     1501 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     1561 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     1621 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     1681 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     1741 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1801 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1861 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1921 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1981 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2041 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2101 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2161 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2221 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2281 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2341 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2401 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2461 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2521 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     2581 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2641 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2701 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2761 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2821 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2881 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2941 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     3001 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     3061 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     3121 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
     3181 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3241 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     3301 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     3361 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     3421 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     3481 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3541 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3601 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     3661 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     3721 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3781 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3841 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtactca
     3901 gatctcgagc tcaagcttat gggggagccg gctggggttg ccggcaccat ggagtcacct
     3961 tttagcccgg gactctttca caggctggat gaagattggg attctgctct ctttgctgaa
     4021 ctcggttatt tcacagacac tgatgagctg caattggaag cagcaaatga gacgtatgaa
     4081 aacaattttg ataatcttga ttttgatttg gatttgttac cttgggagtc agacatttgg
     4141 gacatcaaca accaaatctg tacagttaaa gatattaagg cagaacccca gccactttct
     4201 ccagcctcct caagttattc agtctcatct cctcggtcag tggactctta ttcttcaact
     4261 cagcatgttc ctgaggagtt ggatttgtct tctagttctc agatgtctcc cctttcctta
     4321 tatggtgaaa actctaatag tctctcttca ccggagccac tgaaggaaga taagcctgtc
     4381 actggttcta ggaacaagac tgaaaatgga ctgactccaa agaaaaaaat tcaggtgaat
     4441 tcaaaacctt caattcagcc caagccttta ttgcttccag cagcacccaa gactcaaaca
     4501 aactccagtg ttccagcaaa aaccatcatt attcagacag taccaacgct tatgccattg
     4561 gcaaagcagc aaccaattat cagtttacaa cctgcaccca ctaaaggcca gacggttttg
     4621 ctgtctcagc ctactgtggt acaacttcaa gcacctggag ttctgccctc tgctcagcca
     4681 gtccttgctg ttgctggggg agtcacacag ctccctaatc acgtggtgaa tgtggtacca
     4741 gccccttcag cgaatagccc agtgaatgga aaactttccg tgactaaacc tgtcctacaa
     4801 agtaccatga gaaatgtcgg ttcagatatt gctgtgctaa ggagacagca acgtataata
     4861 aaaaatcgag aatccgcttg tcagtctcgc aagaagaaga aagaatatat gctagggtta
     4921 gaggcgagat taaaggctgc cctctcagaa aacgagcaac tgaagaaaga aaatggaaca
     4981 ctgaagcggc agctggatga agttgtgtca gagaaccaga ggcttaaagt ccctagtcca
     5041 aagcgaagag ttgtctgtgt gatgatagta ttggcattta taatactgaa ctatggacct
     5101 atgagcatgt tggaacagga ttccaggaga atgaatccta gtgtgagccc tgcaaatcaa
     5161 aggaggcacc ttctaggatt ttctgctaaa gaggcacagg acacatcaga tggtattatc
     5221 cagaaaaaca gctacagata tgatcattct gtttcaaatg acaaagccct gatggtgcta
     5281 actgaagaac cattgcttta cattccccca cctccttgtc agcccctaat taatacaaca
     5341 gagtctctca ggttaaatca tgaacttcga ggatgggttc atagacatga agtagaaagg
     5401 accaagtcaa gaagaatgac aaataatcaa cagaaaaccc gtattcttca gggtgctctg
     5461 gaacagggct caaattctca gctgatggct gttcaataca cagaaaccac tagtagtatc
     5521 agcaggaact cagggagtga gctacaagtg tattatgctt cacccagaag ttatcaagac
     5581 ttttttgaag ccatccgcag aaggggagac acattttatg ttgtgtcatt tcgaagggat
     5641 cacctgctgt taccagctac cacccataac aagaccacaa gaccaaaaat gtcaattgtg
     5701 ttaccagcaa taaacataaa tgagaatgtg atcaatgggc aggactacga agtgatgatg
     5761 cagattgact gtcaggtgat ggacaccagg atcctccata tcaaaagttc gtcggttcct
     5821 ccttacctcc gagatcagca gaggaatcaa accaacacct tctttggctc ccctcccgca
     5881 gccacagagg caacccacgt tgtcagcacc atccctgagt cattacaata ggtcgacggt
     5941 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     6001 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     6061 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     6121 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6181 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     6241 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6301 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6361 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6421 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6481 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6541 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6601 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6661 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6721 aatacattc
//
