LOCUS       sequence-9483-ex        6254 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 6254)
  AUTHORS   Merksamer PI, Trusina A, Papa FR
  TITLE     Real-time redox measurements during endoplasmic reticulum stress 
            reveal interlinked protein folding functions.
  JOURNAL   Cell. 2008 Nov 28. 135(5):933-47.
  PUBMED    19026441
REFERENCE   2  (bases 1 to 6254)
  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: "Cell. 2008 
            Nov 28. 135(5):933-47."
COMMENT     Sequence Label: pPM47 (UPR-RFP CEN/ARS URA3)
FEATURES             Location/Qualifiers
     source          1..6254
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..410
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             411..1214
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1343..1798)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1480..1501)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1692..1711
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1928..1950
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1942..1959
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1943..1959
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2457..3167
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2602..2620)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2870..2889
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3178..3425
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3194..3212)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(3434..3450)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3435..3451)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3480..3498)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3519..3535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3519..3535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3532..3554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3543..3559
                     /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(3567..3573,3574..3591,3592..3597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3750..3767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3921..4509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4680..5471,5472..5540))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5303..5322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5541..5645)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5682..6185
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc
      241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
      301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
      361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag
      421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
      481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
      541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
      601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
      661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc
      721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
      781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
      841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
      901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
      961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt
     1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta
     1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg
     1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta
     1981 tagggcgaat tggagctcca ccgcggccat tattatcatg acattaacct ataaaaatag
     2041 gcgtatcacg aggccctttc gtcttcaaga attgatctgt cgacaggaac tggacagcgt
     2101 gtcgaaaaag ctcgacagga actggacagc gtgtcgaaaa agctcgacag gaactggaca
     2161 gcgtgtcgaa aaagctcgac aggaactgga cagcgtgtcg aaaaagctcg agcagatccg
     2221 ccaggcgtgt atatatagcg tggatggcca ggcaacttta gtgctgacac atacaggcat
     2281 atatatatgt gtgcgacgac acatgatcat atggcatgca tgtgctctgt atgtatataa
     2341 aactcttgtt ttcttctttt ctctaaatat tctttcctta tacattaggt cctttgtagc
     2401 ataaattact atacttctat agacacgcaa acacaaatac acacactaaa ttaataatgg
     2461 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     2521 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2581 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     2641 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     2701 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2761 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     2821 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     2881 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     2941 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3001 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     3061 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     3121 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaatct agagtcatca
     3181 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
     3241 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
     3301 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
     3361 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
     3421 tttgcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccag cttttgttcc
     3481 ctttagtgag ggttaattcc gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3541 aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa gtgtaaagcc
     3601 tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3901 aaggccgcgt tgctggcgtt tttccatagg ctcggccccc ctgacgagca tcacaaaaat
     3961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgttcccc
     4021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4081 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     4141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4321 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4441 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4621 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4741 gttgcctgac tgcccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4861 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg aaaaaaagcg
     5101 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5341 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5641 ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc tataaaaata
     5701 attataattt aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa
     5761 attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa
     5821 caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat
     5881 cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta
     5941 atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg
     6001 ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc
     6061 taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga
     6121 gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag
     6181 cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6241 gaggcccttt cgtc
//
