LOCUS       sequence-148115-        8047 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 8047)
  AUTHORS   Wang Q, Mora-Jensen H, Weniger MA, Perez-Galan P, Wolford C, Hai T, 
            Ron D, Chen W, Trenkle W, Wiestner A, Ye Y
  TITLE     ERAD inhibitors integrate ER stress with an epigenetic mechanism to 
            activate BH3-only protein NOXA in cancer cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Feb 17. 106(7):2200-5.
  PUBMED    19164757
REFERENCE   2  (bases 1 to 8047)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Feb 17. 106(7):2200-5."
COMMENT     Sequence Label: pRK-ATF4
FEATURES             Location/Qualifiers
     source          1..8047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(91..96,97..114,115..121)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        198..577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        578..781
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     732..752
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     778..802
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1012..1030
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1043..1062
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1102..1111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1111..1134
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1947..1964
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     promoter        complement(2234..2252)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2272..2289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2278..2502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2548..2976
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2990..3319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3170..3305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3386..4180
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3440..3459)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4050..4069
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4354..4475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4531..4540
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4544..4678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4747..5104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4747..4938
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      4955..5090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5124..5141)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5124..5140)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5133..5155)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5353..5808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5825..5844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5944..5966
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6004..6022)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6090..6194
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6195..6263,6264..7055)
                     /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(6413..6432)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7226..7814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7715..7734
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7968..7985
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
      181 ttaattcgag ctcgcccgac attgattatt gactagttat taatagtaat caattacggg
      241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      781 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      841 aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa cgcggattcc
      901 ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccaccc ccttggcttc
      961 gttagaacgc ggctacaatt aatacataac cttatgtatc atacacatac gatttaggtg
     1021 acactataga ataacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa
     1081 ctgcacctcg gttctaagct tgccgccatg gactacaagg acgacgatga caaggggtcg
     1141 accatgaccg aaatgagctt cctgagcagc gaggtgttgg tgggggactt gatgtccccc
     1201 ttcgaccagt cgggtttggg ggctgaagaa agcctaggtc tcttagatga ttacctggag
     1261 gtggccaagc acttcaaacc tcatgggttc tccagcgaca aggctaaggc gggctcctcc
     1321 gaatggctgg ctgtggatgg gttggtcagt ccctccaaca acagcaagga ggatgccttc
     1381 tccgggacag attggatgtt ggagaaaatg gatttgaagg agttcgactt ggatgccctg
     1441 ttgggtatag atgacctgga aaccatgcca gatgaccttc tgaccacgtt ggatgacact
     1501 tgtgatctct ttgcccccct agtccaggag actaataagc agccccccca gacggtgaac
     1561 ccaattggcc atctcccaga aagtttaaca aaacccgacc aggttgcccc cttcaccttc
     1621 ttacaacctc ttcccctttc cccaggggtc ctgtcctcca ctccagatca ttcctttagt
     1681 ttagagctgg gcagtgaagt ggatatcact gaaggagata ggaagccaga ctacactgct
     1741 tacgttgcca tgatccctca gtgcataaag gaggaagaca ccccttcaga taatgatagt
     1801 ggcatctgta tgagcccaga gtcctatctg gggtctcctc agcacagccc ctctaccagg
     1861 ggctctccaa ataggagcct cccatctcca ggtgttctct gtgggtctgc ccgtcccaaa
     1921 ccttacgatc ctcctggaga gaagatggta gcagcaaaag taaagggtga gaaactggat
     1981 aagaagctga aaaaaatgga gcaaaacaag acagcagcca ctaggtaccg ccagaagaag
     2041 agggcggagc aggaggctct tactggtgag tgcaaagagc tggaaaagaa gaacgaggct
     2101 ctaaaagaga gggcggattc cctggccaag gagatccagt acctgaaaga tttgatagaa
     2161 gaggtccgca aggcaagggg gaagaaaagg gtcccctagg cggccgctcg agcatgcatc
     2221 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2521 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     2941 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3301 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3361 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3421 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3481 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3541 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3601 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3661 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3721 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3781 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3841 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3901 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3961 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4021 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4081 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4141 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4201 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4261 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4321 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgaggatc
     4501 cccgggtacc gagctcgaat tcaatcgatg gccgccatgg cccaacttgt ttattgcagc
     4561 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4621 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcgg
     4681 gaattaattc ggcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
     4741 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4801 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4861 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4921 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4981 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5041 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5101 caaaaagctg ttaacagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5161 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5221 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5281 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5341 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5401 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5641 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5701 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5821 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5881 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5941 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6421 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6721 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8041 attaatg
//
