LOCUS       sequence-199802-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  AUTHORS   Harding HP, Zhang Y, Ron D
  TITLE     Protein translation and folding are coupled by an 
            endoplasmic-reticulum-resident kinase.
  JOURNAL   Nature. 1999 Jan 21. 397(6716):271-4.
  PUBMED    9930704
REFERENCE   2  (bases 1 to 8349)
  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: "Nature. 
            1999 Jan 21. 397(6716):271-4."
COMMENT     Sequence Label: PERK 2: PERK.K618A.9E10.pCDNA
FEATURES             Location/Qualifiers
     source          1..8349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3372..3401
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3432..3450)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3433..3450)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          3534..3599
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3729..3749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4021..4155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4058..4077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4112..4131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5006..5141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5636..5655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(5768..6226)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5905..5926)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6120..6139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(6350..6368)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6436..6540
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6541..6609,6610..7401)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6759..6778)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        7474..7853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7854..8057
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8008..8028
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8102..8121
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8102..8120
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gtctccatgc ccgcgcgtgg ggcgggccgc tgatggagcg cgccacccgg cccgggccgc
       61 gcgcgctgct gctgcttctg ttcctgctgc tgggctgcgc ggcggggatc tcggcggtcg
      121 cgcccgcccg cagtttgctt gctcccgcgt cggagacagt gtttggctta ggggcagcgg
      181 ccgccccgac ttcggccgcg cgggtgcctg cggtggcaac ggccgaagtg accgtggagg
      241 acgccgaggc attgccggct gccgctggcg aaccggagtc acgcgcgacg gagcccgatg
      301 acgacgtgga actgcggcct cgcggcaggt ccttggtaat catcagcact ttagatggac
      361 gaatcgctgc actggatgcc gagaatgatg ggaaaaagca gtgggatttg gacgtggggt
      421 ctggttcctt ggtttcatct agcctcagca agccagaggt gtttgggaac aagatgatca
      481 tcccctccct ggatggagac ctcttccagt gggaccggga ccgagagagc atggaggccg
      541 tccccttcac ggtggagtcc ctgctcgaat cttcctacaa gtttggagat gatgttgttc
      601 tggttggagg gaaatctctg actacatacg gactcagtgc ttacagtgga aagctgaggt
      661 atatctgttc tgccttggga tgtcgccgat gggatagtga tgaaatggaa gaagaggaag
      721 acatcttgct tctgcagcgt acgcagaaga ctgtgcgagc tgtcgggcct cgaagcggca
      781 gtgagaagtg gaatttcagt gttggccact ttgaacttcg gtatattcca gacatggaaa
      841 ctagagccgg attcattgaa agcaccttta aaccgggtgg aaacaaagaa gactctaaaa
      901 ttatttcaga tgtggaagaa caagaagcca ccatgctgga cacagtgata aaagtttccg
      961 ttgctgattg gaaggtcatg gcgtttagta ggaagggagg ccgcctggaa tgggagtacc
     1021 agttttgtac tcccatcgcg tccgcctggc tggtgaggga tggcaaggtc atccccatca
     1081 gcctgtttga tgatacaagt tacacagcca gcgaagaagc cttgggagac gaagaagaca
     1141 ttgtagaggc tgctcgggga gccacagaga acagcgtgta cttagggatg tacagaggcc
     1201 agctgtacct gcagtcgtcc gtcagggtct cagaaaagtt ccctacaagc ccaaaggcct
     1261 tggagtctgt aaatggcgaa aatgcaatta ttcctctgcc gacgatcaaa tggaagccct
     1321 taatccattc tccttctagg actcctgtct tggttgggtc tgatgaattt gacaaatgtc
     1381 taagtaatga taagtattcc cacgaagaat acagtaatgg tgcactttca atcctccagt
     1441 atccatacga taacggttac tatctgccat actacaagag agaaaggaat aagcggagca
     1501 cgcagatcac agtcaggttc ctggacagcc cccactacag caagaacatc cgcaagaagg
     1561 accctatcct cctgctgcac tggtggaagg agatattcgg gacgatcctg ctttgcatcg
     1621 tagccacgac cttcatcgtg cgcaggcttt tccatcctca gccccacagg cagcggaagg
     1681 agtctgaaac tcagtgccag actgaaagta aatacgactc cgtgagtgcc gatgtcagtg
     1741 acaacagctg gaatgacatg aagtactcag gatacgtatc ccgatatcta acagattttg
     1801 agccaattca gtgcatgggt cgtggtggct ttggcgttgt ctttgaagct aaaaacaaag
     1861 tagatgactg caattacgct atcgcgagga tccggctccc caacagggag ttggcacggg
     1921 agaaggtaat gcgggaagtt aaagccttgg ctaagctgga acacccaggc attgtgaggt
     1981 atttcaacgc ctggctggaa accccaccag agaagtggca agaagagatg gatgagatct
     2041 ggctcaaaga cgaaagcaca gactggccgc tcagctcccc tagcccgatg gatgccccat
     2101 ctgttaagat ccgaaggatg gatcctttct ctacaaaaga gcagatcgaa gtcatagctc
     2161 cttctcctga aagaagtcgg tctttctcgg tgggcatttc ctgtggccag acaagctcat
     2221 cggagagcca gttctctccc ctggagttct cagggacaga ctgcggagac aacagtgact
     2281 cagcggacgc agcctacaac ctccaggaca gttgcctgac ggactgcgag gacgtggaag
     2341 atggcaccgt ggacggcaat gacgagggac actcctttga actttgtccg tccgaagctt
     2401 ctccctatac ccggtcaagg gaaggaacgt cctcctccat agtgtttgag gactctggct
     2461 gcggcaacgc gtccagtaag gaggagccca gagggaaccg gctgcatgat ggcaaccatt
     2521 atgttaataa gctaactgat ctcaagtgct ccagcagcag gtcttcttca gaagccacca
     2581 ccttgtctac ctcccctacc aggccaacca ctctaagctt ggatttcacc aagaacactg
     2641 tgggccagct ccagcccagc tcccccaagg tgtatctgta cattcagatg cagctgtgca
     2701 ggaaggagaa cctcaaagac tggatgaacc ggcgctgcag cttggaggac cgggagcacg
     2761 gcgtgtgcct gcacatcttc ctgcagatcg cagaggcagt ggagttcctg cacagcaagg
     2821 gactcatgca cagggacctc aagccttcca acatattctt cacaatggat gatgtggtca
     2881 aggttgggga ctttggactg gtgactgcta tggaccaaga tgaagaagag cagactgtac
     2941 tgactccaat gccagcctat gctacgcaca cgggacaagt agggaccaag ctatacatga
     3001 gcccagagca gattcatgga aacaactact cccataaagt ggacatcttc tctttaggct
     3061 tgattctgtt tgaactcctc tacccattca gcacccagat ggaacgagtc cggattttaa
     3121 ctgatgtcag aaatctcaag tttcctctac tgttcactca gaaatatccc caagagcata
     3181 tgatggttca agacatgctc tctccatccc ccacggagcg gcctgaagcc acagacatca
     3241 ttgaaaatgc catatttgag aacttggagt ttcccgggaa aacggttctg agacagcggt
     3301 cccgctccat gagttcatct ggaacaaaac attccagaca gcccagctgc tcgtacacgc
     3361 cactgcctgg cgagcagaag ctgatctctg aggaggatct gtgactcgag catgcatcta
     3421 gagggcccta ttctatagtg tcacctaaat gctagaggat ctttgtgaag gaaccttact
     3481 tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata
     3541 taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
     3601 ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc
     3661 tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt
     3721 ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc
     3781 taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca
     3841 ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat ttgatgtata
     3901 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3961 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     4021 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4081 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4141 tatcatgtct ggatcgatcc cgccatggta tcaacgccat atttctattt acagtaggga
     4201 cctcttcgtt gtgtaggtac cgctgtattc ctagggaaat agtagaggca ccttgaactg
     4261 tctgcatcag ccatatagcc cccgctgttc gatttacaaa cacaggcaca gtactgacaa
     4321 acccatacac ctcctctgaa atacccatag ttgctagggc tgtctccgaa ctcattacac
     4381 cctccaaagt cagagctgta atttcgccat caagggcagc gagggcttct ccagataaaa
     4441 tagcttctgc cgagagtccc gtaagggtag acacttcagc taatccctcg atgaggtcta
     4501 ctagaatagt cagtgcggct cccattttga aaattcactt acttgatcag cttcagaaga
     4561 tggcggaggg cctccaacac agtaattttc ctcccgactc ttaaaataga aaatgtcaag
     4621 tcagttaagc aggaagtgac taactgaccg cagctggccg tgcgacatcc tcttttaatt
     4681 agttgctagg caactgccct ccagagggca gtgtggtttt gcaagaggaa gcaaaaagcc
     4741 tctccaccca ggcctagaat gtttccaccc aatcattact atgacaacag ctgttttttt
     4801 tagtattaag cagaggccgg gggcccctgg cctccgctta ctctggagaa aaagaagaga
     4861 ggcattgtag aggcttccag aggcaacttg tcaaaacagg actgcttcta tttctgtcac
     4921 actgtctggc cctgtcacaa ggtccagcac ctccataccc cctttaataa gcagtttggg
     4981 aacgggtgcg ggtcttactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     5041 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     5101 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5161 aattcggcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5281 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5341 caccgcctac atacctcgct ctgctgaagc cagttaccag tggctgctgc cagtggcgat
     5401 aagtcgtgtc ttaccgggtt ggactcaaga gatagttacc ggataaggcg cagcggtcgg
     5461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca agctagcttc
     5761 tagctagaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5821 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5881 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5941 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggccgccc actacgtgaa
     6001 ccatcaccca aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6061 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     6121 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     6181 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtacta tggttgcttt
     6241 gacgagcacg tataacgtgc tttcctcgtt agaatcagag cgggagctaa acaggaggcc
     6301 gattaaaggg attttagaca ggaacggtac gccagctgga cccgaaaggg cctcgtgata
     6361 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6421 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6481 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6541 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6601 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6661 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6721 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6781 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6841 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6901 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6961 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     7021 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     7081 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     7141 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     7201 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     7261 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     7321 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7381 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7441 ttaaaacttc atttttaatt tctctagcgc gttgacattg attattgact agttattaat
     7501 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     7561 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     7621 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     7681 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     7741 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     7801 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     7861 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     7921 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     7981 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     8041 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     8101 ttaatacgac tcactatagg gagacccaag cttggtaccg agctcggatc cactagtaac
     8161 ggccgccagt gtgctggaat tcggcacgag cgatgtctgc acaaggctgt cactcaggtg
     8221 gcagtggctg agacgtggcc gggcagctct gctgctgcgg cgcgaagtcg agaggcggcg
     8281 gggtccgtgg cgcgcgctcg cagtgctccg aggctccgag cggcgagacg ggcgggcgcc
     8341 gacggcagg
//
