LOCUS       sequence-202548-        7069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Gga2 with Gga2-msGFPx3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7069)
  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 7069)
  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-GGA2-msGFPx3
FEATURES             Location/Qualifiers
     source          1..7069
                     /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(3567..3584)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3567..3583)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3576..3598)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3792..3811)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3911..3933
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4051..4854)
                     /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(4855..5070)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5077..5095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5163..5267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5268..5336,5337..6128)
                     /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(5486..5505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6299..6887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6788..6807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7041..7058
                     /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 atgctccgtt acaacaatct tctggtgctt taacgaatga aatcaactta atcgatttca
      301 atgatttgga cgaagcgccc tctcaaggca acaacaatac caacggcacg ggtacaccag
      361 cagcagcgga aacatctgtc aatgacttat tgggcgattt gaccgattta tctatttcga
      421 acccttcaac cgctaaccaa gcatcctttg gtctgggagg cgatatcgtg ttggggtcct
      481 ctcaacccgc accaccagtt actactacta acaactcgaa caacacttta gatcttttgg
      541 gactttcaac tcctcaatcg ccaactaatt cccaagcagt aaacagtagt gggttcgatc
      601 ttttgatggg atttaatcca acaacaggta ctactactgc gcccgcaaga accctagtca
      661 atcagtctcc taacttgaag attgagttcg aaatatctag ggagtcaaac tcagttataa
      721 ggataaaatc gttctttaca aacttaagtt cgtcaccgat ctccaactta gtgttcttat
      781 tagcagtccc taagtcaatg tctttgaaat tgcaacccca atcgagtaac tttatgattg
      841 gcaacgctaa agatggtatc tctcaagaag gtacaattga aaatgcgcca gctaaccctt
      901 caaaggcttt gaaagtcaag tggaaggtca actattctgt caactccacc caagctgaag
      961 aaactgctgt ttttacgtta cctaatgtaa cggatgatcc aggaagttcg gggtccagcg
     1021 gttcttcagg cgctagcatg gatagcactg agagcctgtt caccggggtg gtgcccatcc
     1081 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg
     1141 gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1201 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1261 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1321 agcgcaccat caccttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg
     1381 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1441 acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat atcaccgccg
     1501 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg gaggacggca
     1561 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1621 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     1681 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcagctccg
     1741 gctccagcgg gtctagcatg gatagcactg agagcctgtt caccggggtg gtgcccatcc
     1801 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg
     1861 gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1921 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1981 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2041 agcgcaccat caccttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg
     2101 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2161 acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat atcaccgccg
     2221 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg gaggacggca
     2281 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2341 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     2401 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcagctccg
     2461 gctccagcgg gtctagcatg gatagcactg agagcctgtt caccggggtg gtgcccatcc
     2521 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg
     2581 gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2641 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     2701 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2761 agcgcaccat caccttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg
     2821 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2881 acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat atcaccgccg
     2941 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg gaggacggca
     3001 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3061 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
     3121 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcagctccg
     3181 gctccagcgg gtaaagcggc cgctcctctg gcgtttctta tcaatccttt ctcttctcta
     3241 tgtatatttt tagaaggata atacctctat gtagacaatc ttataaagat cttcgcaaca
     3301 atacatattt cacttactaa tatatgttgc gtcttcatgc atcaaaagta gttagcaatg
     3361 ctagatcgtg taaaggaaaa aaaagagaaa tagtataatt cgccatcctc ataaccactg
     3421 aaaaatcgaa gttaaccaaa aggaaagttg aaatggagga agtgttccgt ttttactcga
     3481 actctagaaa tatcttcatt cataaaagcc tttcattaaa accatccacc attgatgacc
     3541 ccaaaagtgg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     3601 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3661 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3721 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3781 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3841 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3901 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3961 agacgaaagg gcgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     4021 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     4081 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     4141 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     4201 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     4261 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     4321 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     4381 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     4441 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     4501 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     4561 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     4621 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     4681 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     4741 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     4801 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     4861 tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     4921 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     4981 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa
     5041 aaaagaataa aaaaaaaatg atgaattgaa aagctggcct cgtgatacgc ctatttttat
     5101 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5161 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5221 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5281 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5341 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5401 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5461 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5521 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5581 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5641 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5701 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5761 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5821 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5881 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5941 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6001 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6061 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6121 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6181 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6241 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6421 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6901 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6961 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7021 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
