LOCUS       sequence-201578-        6458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of RPL13A with an 
            FKBPx4-tagged version..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6458)
  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 6458)
  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: YIplac211-RPL13A-FKBPx4
FEATURES             Location/Qualifiers
     source          1..6458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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     227..249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     246..262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     246..262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(654..656,657..977)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             984..1304
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             984..1304
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1311..1631
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1638..1958
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /note="synthetic gene with alternative codons"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1638..1958
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1965..1991
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2998..3015)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2998..3014)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3007..3029)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3223..3242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3342..3364
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3482..4285)
                     /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"
     promoter        complement(4286..4501)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4508..4526)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4594..4698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4699..4767,4768..5559)
                     /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(4917..4936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5730..6318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6219..6238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
       61 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcacacagga aacagctatg accatgatta cgccaagctt ggccgctggt ttgactgctg
      301 cttatgccag aactatcggt atcgccgttg accacagacg tcaaaacaga aaccaagaaa
      361 tctttgatgc taacgttcaa agattaaagg aataccaatc taagattatt gttttcccaa
      421 gaaatggtaa ggccccagaa gctgaacaag ttttgtctgc tgctgctact ttcccaattg
      481 ctcaaccagc taccgatgtt gaagctagag ctgtccaaga caatggtgaa tctgctttca
      541 gaaccttgag attagccaga tctgagaaga aattcagagg tatcagagaa aagagagcta
      601 gagaaaaggc tgaagctgaa gctgaaaaga agaaatacgc tgcaggtcga cggatgggag
      661 tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct
      721 gcgtggtgca ctacaccggg atgcttgaag atggaaagaa atttgattcc tcccgggaca
      781 gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag
      841 gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct
      901 atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg
      961 agcttctaaa actggaaact agaggcgtgc aggtcgagac catcagcccc ggcgacggcc
     1021 gcacctttcc caagagaggc cagacttgcg tggtccacta caccggcatg ctggaggacg
     1081 gcaagaagtt cgacagcagc cgcgaccgca acaagccctt caagttcatg ctgggcaaac
     1141 aggaagtgat ccgcggctgg gaggaaggcg tggctcagat gagcgtgggg cagcgggcca
     1201 agctgaccat cagccccgac tatgcctacg gcgccaccgg ccaccccggc atcatccccc
     1261 cccacgccac cctggtgttc gacgtggagc tgctgaagct ggagactaga ggagtgcagg
     1321 tggaaaccat ctccccagga gacgggcgca ccttccccaa gcgcggccag acctgcgtgg
     1381 tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg gacagaaaca
     1441 agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa gaaggggttg
     1501 cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat gcctatggtg
     1561 ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat gtggagcttc
     1621 taaaactgga aactagaggc gtgcaggtcg agaccatcag ccccggcgac ggccgcacct
     1681 ttcccaagag aggccagact tgcgtggtcc actacaccgg catgctggag gacggcaaga
     1741 agttcgacag cagccgcgac cgcaacaagc ccttcaagtt catgctgggc aaacaggaag
     1801 tgatccgcgg ctgggaggaa ggcgtggctc agatgagcgt ggggcagcgg gccaagctga
     1861 ccatcagccc cgactatgcc tacggcgcca ccggccaccc cggcatcatc cccccccacg
     1921 ccaccctggt gttcgacgtg gagctgctga agctggagac tagttatccg tacgacgtac
     1981 cagactacgc ataagaaaag tgagcggccg catctaaaaa cttcatcaaa gaatttttca
     2041 tccacaattt ttgtattttc cttctttttt atctgttcaa tttcataaat aatatatatg
     2101 tacaatgtat aaacgggaag ccatcactta taatttatca tttccttaac tttatagtct
     2161 gggtgagaat atggtttaca gtatttttac aagttagcgc agttttcttg atatgatgga
     2221 acgaacatta ttgcattagt agtctgtttg caagcttttc cttcgggcac aacgtggcat
     2281 tatatatagc atagaagaga tgcgaaagat ggtattgaac aaaaagaaag tactgtctaa
     2341 ttgtattaaa agcgactata tagaaaaatt ataaaaaatt ttaaatgtta gaatatgcaa
     2401 tttcttctaa acagtgcggc acttctaatc aaataaaccg aaacccatgt cgtcatcgga
     2461 ttcttcttta gcttcttctt cttccttttc agcttcggct tcaccggctt caccaccagc
     2521 gacaccacca gcgacaccag ctggggcagc agcaccagcg ctgaagttga ccaataagtc
     2581 cttcaagttt tggccgtcca aagccttagc aaaaatatca gcccagatat tttcaactgg
     2641 gacattggca gcgttagtca aagtcaacaa cttttcagaa gagatttcga tttcagagtc
     2701 agccaaaatc aaggcggcgt aagacaaagc ggattcagta gacatttctt cttgctattt
     2761 tagagataat cggtttaagc tgttgtattg gacaaagcgg actatttctt tattcttttg
     2821 atttctttta ctaaaaatta ttcagaacag aagagatcgc aaactttata aagatatttt
     2881 tttcaaatac catgtaaagg cattctttaa aaaatgaaaa aaaaaaaatc attacaaagt
     2941 tgcgtggtat cttcttacga tattcgtacc tcacgatatg tagcagagct cgaattcact
     3001 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     3061 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     3121 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     3181 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     3241 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3301 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3361 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcgggtaat aactgatata
     3421 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     3481 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     3541 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     3601 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     3661 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     3721 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     3781 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     3841 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     3901 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     3961 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     4021 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     4081 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     4141 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     4201 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     4261 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt
     4321 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     4381 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     4441 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     4501 aaagctggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4561 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4621 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4681 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4741 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4801 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4861 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4921 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4981 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5041 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5101 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5161 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5221 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5281 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5341 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5401 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5461 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5521 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5581 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5641 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5701 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5761 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5821 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5881 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5941 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6001 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6061 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6121 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6181 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6241 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6301 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6361 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6421 accgcctttg agtgagctga taccgctcgc cgcagccg
//
