LOCUS       sequence-201574-        5979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid for overexpression of OM45-FKBPx2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5979)
  AUTHORS   Papanikou E, Day KJ, Austin J, Glick BS
  TITLE     COPI selectively drives maturation of the early Golgi.
  JOURNAL   Elife. 2015 Dec 28;4. pii: e13232. doi: 10.7554/eLife.13232.
  PUBMED    26709839
REFERENCE   2  (bases 1 to 5979)
  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: "Elife."; 
            date: "2015-12-28"; volume: "4. pii: e13232. doi"; pages: " 
            10.7554/eLife.13232"
COMMENT     Sequence Label: YIplac204-TC-OM45-FKBPx2
FEATURES             Location/Qualifiers
     source          1..5979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..504
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     CDS             join(1699..1701,1702..2022)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2029..2349
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2029..2349
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2356..2382
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      2413..2660
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2429..2447)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2679..2695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2679..2695)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2692..2714)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2703..2719
                     /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(2727..2733,2734..2751,2752..2757))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2772..2793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2910..2927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3081..3669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3161..3180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3840..4631,4632..4700))
                     /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     4463..4482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4701..4805)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4873..4891
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4893..4994
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4995..5669
                     /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"
     primer_bind     complement(5788..5810)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5910..5929
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaaataccgc atcaggcgga ccttaataca ttcagacact tctgcggtat caccctactt
       61 attcccttcg agattatatc taggaaccca tcaggttggt ggaagattac ccgttctaag
      121 acttttcagc ttcctctatt gatgttacac ctggacaccc cttttctggc atccagtttt
      181 taatcttcag tggcatgtga gattctccga aattaattaa agcaatcaca caattctctc
      241 ggataccacc tcggttgaaa ctgacaggtg gtttgttacg catgctaatg caaaggagcc
      301 tatatacctt tggctcggct gctgtaacag ggaatataaa gggcagcata atttaggagt
      361 ttagtgaact tgcaacattt actattttcc cttcttacgt aaatattttt ctttttaatt
      421 ctaaatcaat ctttttcaat tttttgtttg tattcttttc ttgcttaaat ctataactac
      481 aaaaaacaca tacataaact aaaaatgtca tcaagaataa ttgtcggcag tgcagcattg
      541 gcagctgcca tcacagctag catcatggtc agagaacaga aggccaaggg tcagagaaga
      601 gagggcaacg tctccgctta ctacaacggc caggagtacg gcagttcagc acccgcacag
      661 ttgggaaagc tacataacat aaagcaaggt ataaaggaag atgccttgtc gttaaaagac
      721 gcgcttctgg gcgtatctca aaaggctagg gaagaggctc caaaggtaac taagcgtgtg
      781 atatcaccag aagaggatgc tcagacacgc aagcagctag gccaaaaagc caaggattct
      841 tcctcgcaaa gcatcttcaa ttgggggttt agtgaggctg aaagaaggaa agccatagcc
      901 atcggggaat ttgatactgc taagaagcgt ttcgaagagg cagtggatcg taatgagaag
      961 gagctcttgt ccaccgtgat gagagagaag aaggccgctc tggacagagc atccattgag
     1021 tacgaaaggt acgggagagc cagagacttt aatgagcttt cggacaagct agaccaacag
     1081 gaaaggaaca gtaatccttt gaaacgcctg ttgaagaata acacgggtga cgctaatact
     1141 gaagaagccg ctgcaagaag tgtccaaggc tggggtgata cggcacagga gtttggtaga
     1201 gaagagttgg aggaagccaa gagaaatgct tcttcagagc caagcgaggc gcaaaaacgt
     1261 cttgacgagc tgaagaagat caaggaaaag ggctggtttg gttacaacaa aggggagcaa
     1321 agcgagcaac agattgctga aagggtagcc agaggtttag aaggatgggg tgaaacagcc
     1381 gctcaacttt ccaaggacga aatggacgat ttaagatgga attatgagaa ttcaaagaaa
     1441 caactggaca agaacgtgtc cgatgccatg gactcgttgt ctaaggcgaa ggaggacttg
     1501 aaacagtacg gcagccactg gtggtctgga tggacttcaa aggtcgacaa tgacaagcag
     1561 gctttaaaag atgaggccca aaagaagtac gatgaagcgt tgaaaaagta cgatgaagcc
     1621 aagaacaaat tcaaagaatg gaatgataag ggtgatggta aattctggag ctcgaaaaag
     1681 gaggtacccg gtcgacggat gggagtgcag gtggaaacca tctccccagg agacgggcgc
     1741 accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga
     1801 aagaaatttg attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag
     1861 gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa
     1921 ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca
     1981 catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaactagagg cgtgcaggtc
     2041 gagaccatca gccccggcga cggccgcacc tttcccaaga gaggccagac ttgcgtggtc
     2101 cactacaccg gcatgctgga ggacggcaag aagttcgaca gcagccgcga ccgcaacaag
     2161 cccttcaagt tcatgctggg caaacaggaa gtgatccgcg gctgggagga aggcgtggct
     2221 cagatgagcg tggggcagcg ggccaagctg accatcagcc ccgactatgc ctacggcgcc
     2281 accggccacc ccggcatcat ccccccccac gccaccctgg tgttcgacgt ggagctgctg
     2341 aagctggaga ctagttatcc gtacgacgta ccagactacg catagttggt acccggggat
     2401 cctctagagt catcatgtaa ttagttatgt cacgcttaca ttcacgccct ccccccacat
     2461 ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct atttattttt
     2521 ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     2581 acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag gttttgggac
     2641 gctcgaaggc tttaatttgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     2701 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2761 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2821 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2881 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2941 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3001 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3061 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3121 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3181 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3241 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3301 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3361 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3421 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3481 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3541 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3601 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3661 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3721 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3781 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3841 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3901 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3961 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4021 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4081 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4141 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4201 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4261 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4321 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4381 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4441 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4501 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4561 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4621 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4681 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4741 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4801 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4861 ttaacctata aaaataggcg tatcacgagg ccaattcggt cgaaaaaaga aaaggagagg
     4921 gccaagaggg agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa
     4981 aggcagcttg gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag
     5041 tttgcggctt gcagagcaca gaggccgcag aatgtgcact agattccgat gctgacttgc
     5101 tgggtattat atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa
     5161 tttcaagtct tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg
     5221 tgtttcgtaa tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata
     5281 tcgtccaact gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag
     5341 ttattaaaag actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga
     5401 aacctcattc gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg
     5461 attggaactc gatttctgac tgggttggaa ggcaagagag ccccgagagc ttacatttta
     5521 tgttagctgg tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta
     5581 ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag
     5641 caaatttcgt caaaaatgct aagaaatagg ttattactga gtagtattta tttaagtatt
     5701 gtttgtgcac ttgcctgcaa gccttttgaa aagcaagcat aaaagatcgc cctttcgtct
     5761 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     5821 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     5881 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5941 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggag
//
