LOCUS       sequence-210826-        5622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid for expression of ER localized mCherry at TRP1 
            locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  AUTHORS   Costa EA, Subramanian K, Nunnari J, Weissman JS
  TITLE     Defining the physiological role of SRP in protein-targeting 
            efficiency and specificity.
  JOURNAL   Science. 2018 Jan 18. pii: science.aar3607. doi: 
            10.1126/science.aar3607.
  PUBMED    29348368
REFERENCE   2  (bases 1 to 5622)
  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: "Science. 
            2018 Jan 18. pii: science.aar3607. doi: 10.1126/science.aar3607."
COMMENT     Sequence Label: pJW1665
FEATURES             Location/Qualifiers
     source          1..5622
                     /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        185..465
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             466..1140
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1237..1692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1374..1395)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1586..1605
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1822..1844
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1836..1853
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1837..1853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1860..1879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2338..2347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2344..3051
                     /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(2489..2507)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2757..2776
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3093..3111)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     3266..3282
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3266..3282
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3316..3332)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3317..3333)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(3362..3380)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3401..3417)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3401..3417)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3414..3436)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3425..3441
                     /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(3449..3455,3456..3473,3474..3479))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3632..3649)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3803..4391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4562..5353,5354..5422))
                     /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     5185..5204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5423..5527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5595..5613
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accaaacgac attactatat atataatata ggaagcattt aatagacagc atcgtaatat
      241 atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa
      301 atagcttgtc accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga
      361 attaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg
      421 agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat
      481 ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca
      541 gaatgtgctc tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag
      601 agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat
      661 agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt
      721 ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg
      781 caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac
      841 tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat
      901 tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga
      961 aggcaagaga gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat
     1021 gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag
     1081 acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag
     1141 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgcg gtgtgaaata
     1201 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt
     1261 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1321 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1381 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1441 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1501 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1561 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1621 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1681 tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1741 tgcgggcctc ttcgctatta cgccagctgg cgaagggggg atgtgctgca aggcgattaa
     1801 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     1861 aatacgactc actatagggc gaattggagc tccaccgcgg tggcgcaatc acacaattct
     1921 ctcggatacc acctcggttg aaactgacag gtggtttgtt acgcatgcta atgcaaagga
     1981 gcctatatac ctttggctcg gctgctgtaa cagggaatat aaagggcagc ataatttagg
     2041 agtttagtga acttgcaaca tttactattt tcccttctta cgtaaatatt tttcttttta
     2101 attctaaatc aatctttttc aattttttgt ttgtattctt ttcttgctta aatctataac
     2161 tacaaaaaac acatacataa actaaaaatg tttttcaaca gactaagcgc tggcaagctg
     2221 ctggtaccac tctccgtggt cctgtacgcc cttttcgtgg taatattacc tttacagaat
     2281 tctttccact cctccaatgt tttagttaga ggtgccgatg atggggatcc actagtcgcc
     2341 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2401 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2461 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2521 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2581 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2641 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2701 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2761 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2821 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2881 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2941 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3001 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gcacgacgaa
     3061 ttgtaatcta gagtcatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc
     3121 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     3181 ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     3241 ctgtacagac gcgtgtacgc atggctctag aactagtgga tcccccgggc tgcaggaatt
     3301 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc agcttttgtt
     3361 ccctttagtg agggttaatt ccgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3421 gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata aagtgtaaag
     3481 cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca ctgcccgctt
     3541 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3601 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3661 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3721 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     3781 aaaaggccgc gttgctggcg tttttccata ggctcggccc ccctgacgag catcacaaaa
     3841 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgttcc
     3901 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     3961 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     4021 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4081 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4141 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4201 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4261 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4321 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4381 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4441 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4501 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4561 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4621 tagttgcctg actgcccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4681 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     4801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4861 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgaaaaaaag
     4981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5041 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5401 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5521 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     5581 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
