LOCUS       sequence-168520-        8345 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 8345)
  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 8345)
  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 1: PERK.WT.9E10.pCDNA
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     471..490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(603..1061)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(740..761)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     955..974
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1186..1204)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1272..1376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1377..1445,1446..2237)
                     /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(1595..1614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2310..2689
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2690..2893
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2844..2864
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2938..2957
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2938..2956
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6557..6586
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(6617..6635)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6618..6635)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          6719..6784
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6914..6934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7206..7340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7243..7262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7297..7316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8186..8321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
       61 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      121 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      361 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcaagcta gcttctagct
      601 agaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      661 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga
      721 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      781 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc cgcccactac gtgaaccatc
      841 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      901 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      961 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1021 caccacaccc gccgcgctta atgcgccgct acagggcgcg tactatggtt gctttgacga
     1081 gaccgtataa cgtgctttcc tcgttggaat cagagcggga gctaaacagg aggccgatta
     1141 aagggatttt agacaggaac ggtacgccag ctggattacc aaagggcctc gtgatacgcc
     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2281 aacttcattt ttaatttctc tagcgcgttg acattgatta ttgactagtt attaatagta
     2341 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2401 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2461 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
     2521 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2581 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2641 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2701 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2761 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2821 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2881 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     2941 tacgactcac tatagggaga cccaagcttg gtaccgagct cggatccact agtaacggcc
     3001 gccagtgtgc tggaattcgg cacgagcgat gtctgcacaa ggctgtcact caggtggcag
     3061 tggctgagac gtggccgggc agctctgctg ctgcggcgcg aagtcgagag gcggcggggt
     3121 ccgtggcgcg cgctcgcagt gctccgaggc tccgagcggc gagacgggcg ggcgccgacg
     3181 gcagggtctc catgcccgcg cgtggggcgg gccgctgatg gagcgcgcca cccggcccgg
     3241 gccgcgcgcg ctgctgctgc ttctgttcct gctgctgggc tgcgcggcgg ggatctcggc
     3301 ggtcgcgccc gcccgcagtt tgcttgctcc cgcgtcggag acagtgtttg gcttaggggc
     3361 agcggccgcc ccgacttcgg ccgcgcgggt gcctgcggtg gcaacggccg aagtgaccgt
     3421 ggaggacgcc gaggcattgc cggctgccgc tggcgaaccg gagtcacgcg cgacggagcc
     3481 cgatgacgac gtggaactgc ggcctcgcgg caggtccttg gtaatcatca gcactttaga
     3541 tggacgaatc gctgcactgg atgccgagaa tgatgggaaa aagcagtggg atttggacgt
     3601 ggggtctggt tccttggttt catctagcct cagcaagcca gaggtgtttg ggaacaagat
     3661 gatcatcccc tccctggatg gagacctctt ccagtgggac cgggaccgag agagcatgga
     3721 ggccgtcccc ttcacggtgg agtccctgct cgaatcttcc tacaagtttg gagatgatgt
     3781 tgttctggtt ggagggaaat ctctgactac atacggactc agtgcttaca gtggaaagct
     3841 gaggtatatc tgttctgcct tgggatgtcg ccgatgggat agtgatgaaa tggaagaaga
     3901 ggaagacatc ttgcttctgc agcgtacgca gaagactgtg cgagctgtcg ggcctcgaag
     3961 cggcagtgag aagtggaatt tcagtgttgg ccactttgaa cttcggtata ttccagacat
     4021 ggaaactaga gccggattca ttgaaagcac ctttaaaccg ggtggaaaca aagaagactc
     4081 taaaattatt tcagatgtgg aagaacaaga agccaccatg ctggacacag tgataaaagt
     4141 ttccgttgct gattggaagg tcatggcgtt tagtaggaag ggaggccgcc tggaatggga
     4201 gtaccagttt tgtactccca tcgcgtccgc ctggctggtg agggatggca aggtcatccc
     4261 catcagcctg tttgatgata caagttacac agccagcgaa gaagccttgg gagacgaaga
     4321 agacattgta gaggctgctc ggggagccac agagaacagc gtgtacttag ggatgtacag
     4381 aggccagctg tacctgcagt cgtccgtcag ggtctcagaa aagttcccta caagcccaaa
     4441 ggccttggag tctgtaaatg gcgaaaatgc aattattcct ctgccgacga tcaaatggaa
     4501 gcccttaatc cattctcctt ctaggactcc tgtcttggtt gggtctgatg aatttgacaa
     4561 atgtctaagt aatgataagt attcccacga agaatacagt aatggtgcac tttcaatcct
     4621 ccagtatcca tacgataacg gttactatct gccatactac aagagagaaa ggaataagcg
     4681 gagcacgcag atcacagtca ggttcctgga cagcccccac tacagcaaga acatccgcaa
     4741 gaaggaccct atcctcctgc tgcactggtg gaaggagata ttcgggacga tcctgctttg
     4801 catcgtagcc acgaccttca tcgtgcgcag gcttttccat cctcagcccc acaggcagcg
     4861 gaaggagtct gaaactcagt gccagactga aagtaaatac gactccgtga gtgccgatgt
     4921 cagtgacaac agctggaatg acatgaagta ctcaggatac gtatcccgat atctaacaga
     4981 ttttgagcca attcagtgca tgggtcgtgg tggctttggc gttgtctttg aagctaaaaa
     5041 caaagtagat gactgcaatt acgctatcaa gaggatccgg ctccccaaca gggagttggc
     5101 acgggagaag gtaatgcggg aagttaaagc cttggctaag ctggaacacc caggcattgt
     5161 gaggtatttc aacgcctggc tggaaacccc accagagaag tggcaagaag agatggatga
     5221 gatctggctc aaagacgaaa gcacagactg gccgctcagc tcccctagcc cgatggatgc
     5281 cccatctgtt aagatccgaa ggatggatcc tttctctaca aaagagcaga tcgaagtcat
     5341 agctccttct cctgaaagaa gtcggtcttt ctcggtgggc atttcctgtg gccagacaag
     5401 ctcatcggag agccagttct ctcccctgga gttctcaggg acagactgcg gagacaacag
     5461 tgactcagcg gacgcagcct acaacctcca ggacagttgc ctgacggact gcgaggacgt
     5521 ggaagatggc accgtggacg gcaatgacga gggacactcc tttgaacttt gtccgtccga
     5581 agcttctccc tatacccggt caagggaagg aacgtcctcc tccatagtgt ttgaggactc
     5641 tggctgcggc aacgcgtcca gtaaggagga gcccagaggg aaccggctgc atgatggcaa
     5701 ccattatgtt aataagctaa ctgatctcaa gtgctccagc agcaggtctt cttcagaagc
     5761 caccaccttg tctacctccc ctaccaggcc aaccactcta agcttggatt tcaccaagaa
     5821 cactgtgggc cagctccagc ccagctcccc caaggtgtat ctgtacattc agatgcagct
     5881 gtgcaggaag gagaacctca aagactggat gaaccggcgc tgcagcttgg aggaccggga
     5941 gcacggcgtg tgcctgcaca tcttcctgca gatcgcagag gcagtggagt tcctgcacag
     6001 caagggactc atgcacaggg acctcaagcc ttccaacata ttcttcacaa tggatgatgt
     6061 ggtcaaggtt ggggactttg gactggtgac tgctatggac caagatgaag aagagcagac
     6121 tgtactgact ccaatgccag cctatgctac gcacacggga caagtaggga ccaagctata
     6181 catgagccca gagcagattc atggaaacaa ctactcccat aaagtggaca tcttctcttt
     6241 aggcttgatt ctgtttgaac tcctctaccc attcagcacc cagatggaac gagtccggat
     6301 tttaactgat gtcagaaatc tcaagtttcc tctactgttc actcagaaat atccccaaga
     6361 gcatatgatg gttcaagaca tgctctctcc atcccccacg gagcggcctg aagccacaga
     6421 catcattgaa aatgccatat ttgagaactt ggagtttccc gggaaaacgg ttctgagaca
     6481 gcggtcccgc tccatgagtt catctggaac aaaacattcc agacagccca gctgctcgta
     6541 cacgccactg cctggcgagc agaagctgat ctctgaggag gatctgtgac tcgagcatgc
     6601 atctagaggg ccctattcta tagtgtcacc taaatgctag aggatctttg tgaaggaacc
     6661 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     6721 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     6781 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
     6841 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
     6901 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga
     6961 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
     7021 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatatttgat
     7081 gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag gttttacttg
     7141 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     7201 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7261 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7321 tatcttatca tgtctggatc atcccgccat ggtatcaacg ccatatttct atttacagta
     7381 gggacctctt cgttgtgtag gtaccgctgt attcctaggg aaatagtaga ggcaccttga
     7441 actgtctgca tcagccatat agcccccgct gttcgactta caaacacagg cacagtactg
     7501 acaaacccat acacctcctc tgaaataccc atagttgcta gggctgtctc cgaactcatt
     7561 acaccctcca aagtcagagc tgtaatttcg ccatcaaggg cagcgagggc ttctccagat
     7621 aaaatagctt ctgccgagag tcccgtaagg gtagacactt cagctaatcc ctcgatgagg
     7681 tctactagaa tagtcagtgc ggctcccatt ttgaaaattc acttacttga tcagcttcag
     7741 aagatggcgg agggcctcca acacagtaat tttcctcccg actcttaaaa tagaaaatgt
     7801 caagtcagtt aagcaggaag tggactaact gacgcagctg gccgtgcgac atcctctttt
     7861 aattagttgc taggcaacgc cctccagagg gcgtgtggtt ttgcaagagg aagcaaaagc
     7921 ctctccaccc aggcctagaa tgtttccacc caatcattac tatgacaaca gctgtttttt
     7981 ttagtattaa gcagaggccg gggacccctg ggcccgctta ctctggagaa aaagaagaga
     8041 ggcattgtag aggcttccag aggcaacttg tcaaaacagg actgcttcta tttctgtcac
     8101 actgtctggc cctgtcacaa ggtccagcac ctccataccc cctttaataa gcagtttggg
     8161 aacgggtgcg ggtcttactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     8221 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     8281 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     8341 aattc
//
