LOCUS       sequence-202433-        9217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Kex2 with Kex2-iGFP-x6..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9217)
  AUTHORS   Day KJ, Casler JC, Glick BS
  TITLE     Budding Yeast Has a Minimal Endomembrane System.
  JOURNAL   Dev Cell. 2018 Jan 8;44(1):56-72.e4. doi: 
            10.1016/j.devcel.2017.12.014. Epub 2018 Jan 8.
  PUBMED    29316441
REFERENCE   2  (bases 1 to 9217)
  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"; doi: 
            "10.1016/j.devcel.2017.12.014"; journalName: "Dev Cell"; date: 
            "2018-01-8- 8"; volume: "44"; issue: "1"; pages: "56-72.e4"
COMMENT     Sequence Label: YIplac211-KEX2-iGFPx6
FEATURES             Location/Qualifiers
     source          1..9217
                     /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"
     primer_bind     complement(5715..5732)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5715..5731)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5724..5746)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5940..5959)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6059..6081
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6199..7002)
                     /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(7003..7218)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7225..7243)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7311..7415
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7416..7484,7485..8276)
                     /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(7634..7653)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8447..9035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8936..8955
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9189..9206
                     /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 atgcacattg gggtgagaac ggcgtaggtg attggaaaat caaggttaag acaacagaaa
      301 atggacacag gattgacttc cacagttgga ggctgaagct ctttggggaa tccattgatt
      361 catctaaaac agaaactttc gtctttggaa acgataaaga ggaggttgaa ccagctgcta
      421 cagaaagtac cgtatcacaa tattctgcca gttcaacttc tatttccatc agcgctactt
      481 ctacatcttc tatctcaatt ggtgtggaaa cgtcggccat tccccaaacg actactgcga
      541 gtaccgatcc tgattctgat ccaaacactc ctaaaaaact ttcctctcct aggcaagcca
      601 tgcattattt tttaacaata tttttgattg gcgccacatt tttggtgtta tacttcatgt
      661 tttttatgaa atcaaggaga aggatcagaa ggtcaagagc ggaaacgtat gaattcgata
      721 tcattgatac agactctgag tacgattcta ctttggacaa tggaacttcc ggaattactg
      781 agcccgaaga ggttgaggac ttcgattttg atttgtccga tgaagaccat cttgcaagtt
      841 tgtcttcatc agaaaacggt gatgctgaac atacaattga tagtgtacta acaaacgaaa
      901 atccatttag tgaccctata aagcaaaagt tcccaaatga cgccaacgca gaatctgctt
      961 ccaataaatt acaggaatta cagcctgatg ttcctccatc ttccggacga tcgacggatc
     1021 caggaagttc ggggtccagc ggttcttcag gcgctagcat ggatagcact gagagccttt
     1081 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     1141 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     1201 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     1261 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     1321 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     1381 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     1441 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     1501 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     1561 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     1621 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     1681 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     1741 ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact gagagccttt
     1801 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     1861 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     1921 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     1981 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     2041 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     2101 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     2161 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     2221 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     2281 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     2341 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     2401 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     2461 ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact gagagccttt
     2521 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     2581 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     2641 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     2701 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     2761 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     2821 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     2881 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     2941 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     3001 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     3061 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     3121 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     3181 ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact gagagccttt
     3241 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     3301 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     3361 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     3421 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     3481 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     3541 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     3601 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     3661 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     3721 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     3781 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     3841 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     3901 ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact gagagccttt
     3961 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     4021 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     4081 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     4141 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     4201 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     4261 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     4321 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     4381 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     4441 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     4501 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     4561 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     4621 ggattacaca tggcagctcc ggctccagcg ggtctagcat ggatagcact gagagccttt
     4681 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     4741 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     4801 gcactactgg aaaactacct gttccttggc caacacttgt cactacttta acttatggtg
     4861 ttcagtgctt ttcaagatac ccggatcata tgaaacagca tgactttttc aagagtgcca
     4921 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     4981 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     5041 ttgattttaa agaagatgga aacattcttg ggcacaaatt ggaatacaac tataactcac
     5101 acaatgtata catcatggca gacaaacaaa agaatggaat caaagctaac ttcaaaatta
     5161 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     5221 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     5281 cgaaagatcc caacgaaaag agagaccaca tggtccttaa agagtttgta acagctgctg
     5341 ggattacaca tggcagctcc ggctccagcg ggtaaagcgg ccgcttcgat atgtacagaa
     5401 agcttcaaat tacaaaatag catttttttc ttatagatta taatactctc tcatacgtat
     5461 acgtatatgt gtatatgata tataaacaaa cattaatatc ctattccttc cgtttgaaat
     5521 ccctatgatg tactttgcat tgtttgcacc cgcgaataaa atgaaaactc cgaaccgata
     5581 tatcaagcac ataaaagggg agggtccaat taatgcatat ttaagaccac agctgaataa
     5641 ctttaaaacg gcagacaaaa caaaaaatag gtcgaataaa ccttacctgc ctagaaggta
     5701 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5761 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5821 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     5881 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     5941 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6001 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6061 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6121 gggtaataac tgatataatt aaattgaagc tctaatttgt gagtttagta tacatgcatt
     6181 tacttataat acagtttttt agttttgctg gccgcatctt ctcaaatatg cttcccagcc
     6241 tgcttttctg taacgttcac cctctacctt agcatccctt ccctttgcaa atagtcctct
     6301 tccaacaata ataatgtcag atcctgtaga gaccacatca tccacggttc tatactgttg
     6361 acccaatgcg tctcccttgt catctaaacc cacaccgggt gtcataatca accaatcgta
     6421 accttcatct cttccaccca tgtctctttg agcaataaag ccgataacaa aatctttgtc
     6481 gctcttcgca atgtcaacag tacccttagt atattctcca gtagataggg agcccttgca
     6541 tgacaattct gctaacatca aaaggcctct aggttccttt gttacttctt ctgccgcctg
     6601 cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca
     6661 ttctgctatt ctgtatacac ccgcagagta ctgcaatttg actgtattac caatgtcagc
     6721 aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg gataatgcct ttagcggctt
     6781 aactgtgccc tccatggaaa aatcagtcaa gatatccaca tgtgttttta gtaaacaaat
     6841 tttgggacct aatgcttcaa ctaactccag taattccttg gtggtacgaa catccaatga
     6901 agcacacaag tttgtttgct tttcgtgcat gatattaaat agcttggcag caacaggact
     6961 aggatgagta gcagcacgtt ccttatatgt agctttcgac atgatttatc ttcgtttcct
     7021 gcatgttttt gttctgtgca gttgggttaa gaatactggg caatttcatg tttcttcaac
     7081 actacatatg cgtatatata ccaatctaag tctgtgctcc ttccttcgtt cttccttctg
     7141 ttcggagatt accgaatcaa aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa
     7201 aaaaaatgat gaattgaaaa gctggcctcg tgatacgcct atttttatag gttaatgtca
     7261 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     7321 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     7381 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     7441 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     7501 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     7561 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     7621 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     7681 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7741 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7801 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7861 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7921 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7981 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8041 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8101 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8161 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     8221 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     8281 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     8341 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     8401 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     8461 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     8521 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     8581 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     8641 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     8701 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8761 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8821 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8881 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8941 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9001 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9061 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     9121 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9181 ccgagcgcag cgagtcagtg agcgaggaag cggaaga
//
