LOCUS       sequence-233277-        7537 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 7537)
  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 7537)
  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-ATF3
FEATURES             Location/Qualifiers
     source          1..7537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(188..643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     841..863
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     855..872
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     856..872
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(892..1249)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      906..1041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        1058..1249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     polyA_signal    1318..1452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1456..1465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1521..1642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1816..2610)
                     /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(1927..1946)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2537..2556
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2677..3006)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2691..2826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(3020..3448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    3494..3718
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3707..3724
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3744..3762
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(4352..4375)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4375..4384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4424..4443)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(4456..4474)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4684..4708)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4705..4908)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4734..4754)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4909..5288
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(5317..5333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5317..5333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5330..5352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5341..5357
                     /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)."
     promoter        complement(join(5365..5371,5372..5389,5390..5395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5410..5431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5548..5565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5719..6307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5799..5818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6478..7269,7270..7338))
                     /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     7101..7120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7339..7443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7511..7529
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      241 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      301 cccgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      361 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      421 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      481 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      541 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      601 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtactatgg ttgctttgac
      661 gtatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
      721 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
      781 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
      841 cccagtcacg acgttgtaaa acgacggcca gtgccaagct gttaacagct ttttgcaaaa
      901 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
      961 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     1021 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     1081 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     1141 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     1201 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcaga aggtacctaa
     1261 ccaagttcct ctttcagagg ttatttcagg ccatggtgct gcgccgaatt aattcccgat
     1321 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1381 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1441 aataaacaag ttgggccatg gcggccatcg attgaattcg agctcggtac ccggggatcc
     1501 tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1561 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1621 tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga tccccgcgct
     1681 ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg
     1741 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
     1801 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
     1861 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
     1921 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
     1981 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
     2041 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa
     2101 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
     2161 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
     2221 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
     2281 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
     2341 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
     2401 cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt
     2461 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
     2521 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
     2581 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
     2641 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa
     2701 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata
     2761 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     2821 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     2881 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     2941 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     3001 acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac
     3061 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3121 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3181 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3241 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3301 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3361 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3421 gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt
     3481 tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
     3541 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
     3601 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     3661 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
     3721 gaggctgatc agcgagctct agcatttagg tgacactata gaatagggcc ctctagatgc
     3781 atgctcgagc ggccgcttag ctctgcaatg ttccttcttt tatctgttgg ataaagaggt
     3841 ttctctcatc ttctggagtc ctcccattct gagcccggac aatacacgtg ggccgatgaa
     3901 ggttgagcat gtatatcaaa tgctgcttct cgttcttgag ctcctcaatc tgagccttca
     3961 gttcagcatt cacactttcc agcttctccg actctttctg caggcactcc gtcttctcct
     4021 tcttcttgtt tcggcacttt gcagctgcaa tcttatttct ttctcgtcgc ctctttttcc
     4081 tttcatcttc ttcaggggct acctcggctt ttgtgatgga caccccgagg ggtctgtcgc
     4141 tgacagtgac tgattccagc gcagaggaca tccggtggca gaggtgcttg ttctggatgg
     4201 caaacctcag ctcttccttg acaaagggcg tcaggttagc aaaatcctca aacaccagtg
     4261 acccaggagg ggacaggcag gggacgatgg cagaagcact cacttccgag gcagagacct
     4321 ggcctgggtg ttgaagcatc atggtcgacc ccttgtcatc gtcgtccttg tagtccatgg
     4381 cggcaagctt agaaccgagg tgcagttgga cctgggagtg gacacctgtg gagagaaagg
     4441 caaagtggat gttattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat
     4501 gtattaattg tagccgcgtt ctaacgaagc caagggggtg ggcctataga ctctataggc
     4561 ggtacttacg tcactcttgg cacggggaat ccgcgttcca atgcaccgtt cccggccgcg
     4621 gaggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca
     4681 ggcgatctga cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct
     4741 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     4801 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     4861 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     4921 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     4981 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     5041 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     5101 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     5161 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     5221 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     5281 tcaatgtcgg gcgagctcga attaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     5341 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5401 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5461 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6121 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6421 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6721 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6901 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7021 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7141 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7201 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7321 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7381 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7441 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     7501 aacctataaa aataggcgta tcacgaggcc ctttcgt
//
