LOCUS       sequence-202851-        6633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid for overexpression of OM45-FKBPx4..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  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 6633)
  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-FKBPx4
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(239..486)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     452..470
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(517..543)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(550..870)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(550..870)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(877..1197)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(1204..1524)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(1204..1524)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(join(1531..1851,1852..1854))
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     promoter        complement(3049..3479)
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     primer_bind     complement(3603..3622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3722..3744
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3863..4537)
                     /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"
     promoter        complement(4538..4639)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4641..4659)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4727..4831
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4832..4900,4901..5692)
                     /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(5050..5069)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5863..6451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6352..6371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6605..6622
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 aaattaaagc cttcgagcgt cccaaaacct tctcaagcaa ggttttcagt ataatgttac
      301 atgcgtacac gcgtctgtac agaaaaaaaa gaaaaatttg aaatataaat aacgttctta
      361 atactaacat aactataaaa aaataaatag ggacctagac ttcaggttgt ctaactcctt
      421 ccttttcggt tagagcggat gtggggggag ggcgtgaatg taagcgtgac ataactaatt
      481 acatgatgac tctagaggat ccccgggtac caactatgcg tagtctggta cgtcgtacgg
      541 ataactagtc tccagcttca gcagctccac gtcgaacacc agggtggcgt ggggggggat
      601 gatgccgggg tggccggtgg cgccgtaggc atagtcgggg ctgatggtca gcttggcccg
      661 ctgccccacg ctcatctgag ccacgccttc ctcccagccg cggatcactt cctgtttgcc
      721 cagcatgaac ttgaagggct tgttgcggtc gcggctgctg tcgaacttct tgccgtcctc
      781 cagcatgccg gtgtagtgga ccacgcaagt ctggcctctc ttgggaaagg tgcggccgtc
      841 gccggggctg atggtctcga cctgcacgcc tctagtttcc agttttagaa gctccacatc
      901 gaagacgaga gtggcatgtg gtgggatgat gcctgggtgc ccagtggcac cataggcata
      961 atctggagat atagtcagtt tggctctctg acccacactc atctgggcaa ccccttcttc
     1021 ccagcctcgg atcacctcct gcttgcctag cataaactta aagggcttgt ttctgtcccg
     1081 ggaggaatca aatttctttc catcttcaag catcccggtg tagtgcacca cgcaggtctg
     1141 gccgcgcttg gggaaggtgc gcccgtctcc tggggagatg gtttccacct gcactcctct
     1201 agtctccagc ttcagcagct ccacgtcgaa caccagggtg gcgtgggggg ggatgatgcc
     1261 ggggtggccg gtggcgccgt aggcatagtc ggggctgatg gtcagcttgg cccgctgccc
     1321 cacgctcatc tgagccacgc cttcctccca gccgcggatc acttcctgtt tgcccagcat
     1381 gaacttgaag ggcttgttgc ggtcgcggct gctgtcgaac ttcttgccgt cctccagcat
     1441 gccggtgtag tggaccacgc aagtctggcc tctcttggga aaggtgcggc cgtcgccggg
     1501 gctgatggtc tcgacctgca cgcctctagt ttccagtttt agaagctcca catcgaagac
     1561 gagagtggca tgtggtggga tgatgcctgg gtgcccagtg gcaccatagg cataatctgg
     1621 agatatagtc agtttggctc tctgacccac actcatctgg gcaacccctt cttcccagcc
     1681 tcggatcacc tcctgcttgc ctagcataaa cttaaagggc ttgtttctgt cccgggagga
     1741 atcaaatttc tttccatctt caagcatccc ggtgtagtgc accacgcagg tctggccgcg
     1801 cttggggaag gtgcgcccgt ctcctgggga gatggtttcc acctgcactc ccatccgtcg
     1861 accgggtacc tcctttttcg agctccagaa tttaccatca cccttatcat tccattcttt
     1921 gaatttgttc ttggcttcat cgtacttttt caacgcttca tcgtacttct tttgggcctc
     1981 atcttttaaa gcctgcttgt cattgtcgac ctttgaagtc catccagacc accagtggct
     2041 gccgtactgt ttcaagtcct ccttcgcctt agacaacgag tccatggcat cggacacgtt
     2101 cttgtccagt tgtttctttg aattctcata attccatctt aaatcgtcca tttcgtcctt
     2161 ggaaagttga gcggctgttt caccccatcc ttctaaacct ctggctaccc tttcagcaat
     2221 ctgttgctcg ctttgctccc ctttgttgta accaaaccag cccttttcct tgatcttctt
     2281 cagctcgtca agacgttttt gcgcctcgct tggctctgaa gaagcatttc tcttggcttc
     2341 ctccaactct tctctaccaa actcctgtgc cgtatcaccc cagccttgga cacttcttgc
     2401 agcggcttct tcagtattag cgtcacccgt gttattcttc aacaggcgtt tcaaaggatt
     2461 actgttcctt tcctgttggt ctagcttgtc cgaaagctca ttaaagtctc tggctctccc
     2521 gtacctttcg tactcaatgg atgctctgtc cagagcggcc ttcttctctc tcatcacggt
     2581 ggacaagagc tccttctcat tacgatccac tgcctcttcg aaacgcttct tagcagtatc
     2641 aaattccccg atggctatgg ctttccttct ttcagcctca ctaaaccccc aattgaagat
     2701 gctttgcgag gaagaatcct tggctttttg gcctagctgc ttgcgtgtct gagcatcctc
     2761 ttctggtgat atcacacgct tagttacctt tggagcctct tccctagcct tttgagatac
     2821 gcccagaagc gcgtctttta acgacaaggc atcttccttt ataccttgct ttatgttatg
     2881 tagctttccc aactgtgcgg gtgctgaact gccgtactcc tggccgttgt agtaagcgga
     2941 gacgttgccc tctcttctct gacccttggc cttctgttct ctgaccatga tgctagctgt
     3001 gatggcagct gccaatgctg cactgccgac aattattctt gatgacattt ttagtttatg
     3061 tatgtgtttt ttgtagttat agatttaagc aagaaaagaa tacaaacaaa aaattgaaaa
     3121 agattgattt agaattaaaa agaaaaatat ttacgtaaga agggaaaata gtaaatgttg
     3181 caagttcact aaactcctaa attatgctgc cctttatatt ccctgttaca gcagccgagc
     3241 caaaggtata taggctcctt tgcattagca tgcgtaacaa accacctgtc agtttcaacc
     3301 gaggtggtat ccgagagaat tgtgtgattg ctttaattaa tttcggagaa tctcacatgc
     3361 cactgaagat taaaaactgg atgccagaaa aggggtgtcc aggtgtaaca tcaatagagg
     3421 aagctgaaaa gtcttagaac gggtaatctt ccaccaacct gatgggttcc tagatataat
     3481 ctcgaaggga ataagtaggg tgataccgca gaagtgtctg aatgtattaa ggtccgcctg
     3541 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     3601 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3661 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3721 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3781 ggcgatcttt tatgcttgct tttcaaaagg cttgcaggca agtgcacaaa caatacttaa
     3841 ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt
     3901 tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat
     3961 ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta
     4021 agctctcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt
     4081 cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag
     4141 ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac
     4201 cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg atatcaatgc
     4261 cgtaatcatt gaccagagcc aaaacatcct ccttaggttg attacgaaac acgccaacca
     4321 agtatttcgg agtgcctgaa ctatttttat atgcttttac aagacttgaa attttccttg
     4381 caataaccgg gtcaattgtt ctctttctat tgggcacaca tataataccc agcaagtcag
     4441 catcggaatc tagtgcacat tctgcggcct ctgtgctctg caagccgcaa actttcacca
     4501 atggaccaga actacctgtg aaattaataa cagacatact ccaagctgcc tttgtgtgct
     4561 taatcacgta tactcacgtg ctcaatagtc accaatgccc tccctcttgg ccctctcctt
     4621 ttcttttttc gaccgaattg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4681 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4741 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4801 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4861 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4921 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4981 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5041 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5101 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5161 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5221 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5281 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5341 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5401 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5461 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5521 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5581 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5641 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5701 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5761 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5821 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5881 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5941 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6001 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6061 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6121 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6181 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6241 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6301 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6361 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6421 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6481 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6541 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6601 gcgcagcgag tcagtgagcg aggaagcgga aga
//
