LOCUS       sequence-308971-       10184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Sec7 with 
            Sec7-mScarlet2Ix6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10184)
  AUTHORS   Casler JC, Glick BS
  TITLE     A microscopy-based kinetic analysis of yeast vacuolar protein 
            sorting.
  JOURNAL   Elife. 2020 Jun 25;9. pii: 56844. doi: 10.7554/eLife.56844.
  PUBMED    32584255
REFERENCE   2  (bases 1 to 10184)
  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: "2020-06-25"; volume: "9. pii: 56844. doi"; pages: " 
            10.7554/eLife.56844"
COMMENT     Sequence Label: YIplac211-Sec7-mScarlet2Ix6
FEATURES             Location/Qualifiers
     source          1..10184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6842..6858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6842..6858)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6855..6877)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6866..6882
                     /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(6890..6896,6897..6914,6915..6920))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6935..6956
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7073..7090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7244..7832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7324..7343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8003..8794,8795..8863))
                     /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     8626..8645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8864..8968)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(9098..9898)
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQLN"
     promoter        complement(9899..10114)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     10157..10175
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ctattgttgt caaatgtgtt
      421 ttacagcttt tgatgataga gctgctcaac gaattattcg agaacgaaga ttttgcccac
      481 tgtatccctt acaaggaagc cattagaatt acaagattgt tggagaaatc gtacgaattt
      541 tctcgtgact ttaatgaaga ttatgggtta agaacaagac tagtagaggc tcgtgtagtt
      601 gataaaatac caaacctgtt gaaacaagaa acgagtgctg ctgcagttct ccttgacatt
      661 atgtttcagt tgtatctgaa cgatgatgag aagaaggctg atttaataac tcgtctgatc
      721 accatttgca tccaggtcgt agagggttat gtttctttgg atgatagaac catggaacgt
      781 agtatcaatg cctggcgttc agtgatagtt gaaatattac aaggttacta cgaatttgat
      841 gatgaagact ttagactata ctgtcctgct atgtatgctc tggtgattca aatattagat
      901 aagagcgttc caacggagtt gagacatgct ataaaacaat ttctaagcag agttggtgaa
      961 ttataccttt ctacggatcc aactagtatg gatagtacag aagcagttat aaaggaattc
     1021 atgagattca aagtccacat ggaaggtagt atgaacggtc acgaattcga aatagaaggt
     1081 gaaggtgaag gtagaccata tgaaggtact caaacagcta aattgaaggt tactaaaggt
     1141 ggtccattgc cattttcttg ggatatcttg tctccacaat tcatgtacgg ttcaagagct
     1201 tttattaaac atccagcaga tattccagat tactacaagc aatcatttcc agaaggtttt
     1261 aaatgggaaa gagttatgaa cttcgaagat ggtggtgcag ttactgttac acaagatact
     1321 tctttggaag atggtacatt gatctataag gttaaattga gaggtacaaa ttttccacca
     1381 gatggtccag ttatgcaaaa gaaaactatg ggttgggaag catctacaga aagattgtac
     1441 ccagaagatg gtgttttgaa gggtgacatc aagatggcat tgagattgaa ggatggtggt
     1501 agatatttgg ctgatttcaa gactacatac aaggctaaga aaccagttca aatgccaggt
     1561 gcttacaacg ttgatagaaa attggatatc acttctcata acgaagatta cacagttgtt
     1621 gaacaatacg aaagaagtga aggtagacat agtacaggtg gtagtggtgc tagtatggat
     1681 agtacagaag cagttataaa ggaattcatg agattcaaag tccacatgga aggtagtatg
     1741 aacggtcacg aattcgaaat agaaggtgaa ggtgaaggta gaccatatga aggtactcaa
     1801 acagctaaat tgaaggttac taaaggtggt ccattgccat tttcttggga tatcttgtct
     1861 ccacaattca tgtacggttc aagagctttt attaaacatc cagcagatat tccagattac
     1921 tacaagcaat catttccaga aggttttaaa tgggaaagag ttatgaactt cgaagatggt
     1981 ggtgcagtta ctgttacaca agatacttct ttggaagatg gtacattgat ctataaggtt
     2041 aaattgagag gtacaaattt tccaccagat ggtccagtta tgcaaaagaa aactatgggt
     2101 tgggaagcat ctacagaaag attgtaccca gaagatggtg ttttgaaggg tgacatcaag
     2161 atggcattga gattgaagga tggtggtaga tatttggctg atttcaagac tacatacaag
     2221 gctaagaaac cagttcaaat gccaggtgct tacaacgttg atagaaaatt ggatatcact
     2281 tctcataacg aagattacac agttgttgaa caatacgaaa gaagtgaagg tagacatagt
     2341 acaggtggta gtggtgctag tatggatagt acagaagcag ttataaagga attcatgaga
     2401 ttcaaagtcc acatggaagg tagtatgaac ggtcacgaat tcgaaataga aggtgaaggt
     2461 gaaggtagac catatgaagg tactcaaaca gctaaattga aggttactaa aggtggtcca
     2521 ttgccatttt cttgggatat cttgtctcca caattcatgt acggttcaag agcttttatt
     2581 aaacatccag cagatattcc agattactac aagcaatcat ttccagaagg ttttaaatgg
     2641 gaaagagtta tgaacttcga agatggtggt gcagttactg ttacacaaga tacttctttg
     2701 gaagatggta cattgatcta taaggttaaa ttgagaggta caaattttcc accagatggt
     2761 ccagttatgc aaaagaaaac tatgggttgg gaagcatcta cagaaagatt gtacccagaa
     2821 gatggtgttt tgaagggtga catcaagatg gcattgagat tgaaggatgg tggtagatat
     2881 ttggctgatt tcaagactac atacaaggct aagaaaccag ttcaaatgcc aggtgcttac
     2941 aacgttgata gaaaattgga tatcacttct cataacgaag attacacagt tgttgaacaa
     3001 tacgaaagaa gtgaaggtag acatagtaca ggtggtagtg gtgctagtat ggatagtaca
     3061 gaagcagtta taaaggaatt catgagattc aaagtccaca tggaaggtag tatgaacggt
     3121 cacgaattcg aaatagaagg tgaaggtgaa ggtagaccat atgaaggtac tcaaacagct
     3181 aaattgaagg ttactaaagg tggtccattg ccattttctt gggatatctt gtctccacaa
     3241 ttcatgtacg gttcaagagc ttttattaaa catccagcag atattccaga ttactacaag
     3301 caatcatttc cagaaggttt taaatgggaa agagttatga acttcgaaga tggtggtgca
     3361 gttactgtta cacaagatac ttctttggaa gatggtacat tgatctataa ggttaaattg
     3421 agaggtacaa attttccacc agatggtcca gttatgcaaa agaaaactat gggttgggaa
     3481 gcatctacag aaagattgta cccagaagat ggtgttttga agggtgacat caagatggca
     3541 ttgagattga aggatggtgg tagatatttg gctgatttca agactacata caaggctaag
     3601 aaaccagttc aaatgccagg tgcttacaac gttgatagaa aattggatat cacttctcat
     3661 aacgaagatt acacagttgt tgaacaatac gaaagaagtg aaggtagaca tagtacaggt
     3721 ggtagtggtg ctagtatgga tagtacagaa gcagttataa aggaattcat gagattcaaa
     3781 gtccacatgg aaggtagtat gaacggtcac gaattcgaaa tagaaggtga aggtgaaggt
     3841 agaccatatg aaggtactca aacagctaaa ttgaaggtta ctaaaggtgg tccattgcca
     3901 ttttcttggg atatcttgtc tccacaattc atgtacggtt caagagcttt tattaaacat
     3961 ccagcagata ttccagatta ctacaagcaa tcatttccag aaggttttaa atgggaaaga
     4021 gttatgaact tcgaagatgg tggtgcagtt actgttacac aagatacttc tttggaagat
     4081 ggtacattga tctataaggt taaattgaga ggtacaaatt ttccaccaga tggtccagtt
     4141 atgcaaaaga aaactatggg ttgggaagca tctacagaaa gattgtaccc agaagatggt
     4201 gttttgaagg gtgacatcaa gatggcattg agattgaagg atggtggtag atatttggct
     4261 gatttcaaga ctacatacaa ggctaagaaa ccagttcaaa tgccaggtgc ttacaacgtt
     4321 gatagaaaat tggatatcac ttctcataac gaagattaca cagttgttga acaatacgaa
     4381 agaagtgaag gtagacatag tacaggtggt agtggtgcta gtatggatag tacagaagca
     4441 gttataaagg aattcatgag attcaaagtc cacatggaag gtagtatgaa cggtcacgaa
     4501 ttcgaaatag aaggtgaagg tgaaggtaga ccatatgaag gtactcaaac agctaaattg
     4561 aaggttacta aaggtggtcc attgccattt tcttgggata tcttgtctcc acaattcatg
     4621 tacggttcaa gagcttttat taaacatcca gcagatattc cagattacta caagcaatca
     4681 tttccagaag gttttaaatg ggaaagagtt atgaacttcg aagatggtgg tgcagttact
     4741 gttacacaag atacttcttt ggaagatggt acattgatct ataaggttaa attgagaggt
     4801 acaaattttc caccagatgg tccagttatg caaaagaaaa ctatgggttg ggaagcatct
     4861 acagaaagat tgtacccaga agatggtgtt ttgaagggtg acatcaagat ggcattgaga
     4921 ttgaaggatg gtggtagata tttggctgat ttcaagacta catacaaggc taagaaacca
     4981 gttcaaatgc caggtgctta caacgttgat agaaaattgg atatcacttc tcataacgaa
     5041 gattacacag ttgttgaaca atacgaaaga agtgaaggta gacatagtac aggtggtagt
     5101 ggtgctagct aagcggccgg gctttgttga atggtccagc agattaaaat atgcttagtt
     5161 gtagaacaca gaaaaaatga gaaggttgat atacttttac tcatatttac cctgatttaa
     5221 tggcggtgat cctttttaga ttcgtttaat tcataaacaa ggtacatcat atagcaataa
     5281 caaatatgta aactatgtag aaagacttat caagtttagc catcttggtc ctcattcatg
     5341 ggcattttat actaacttca tttttgccga aataagaaag gatgtaatct taacctaaaa
     5401 tatgtaagca attaagggtg cacgtagtta cttgcgaaca aataggaaaa gaaagaaaca
     5461 agcacggata ctgaatgact gagtgtatta ctacacgatt tatctcaacg aatttaagtg
     5521 tggaggacga tcggctgcac gtaggagtca acgttttttt actgtgttca tcacaaaacc
     5581 attaagctga gtcatccata ttgcaaacaa taaggtagca ccgcttccat caaaacacta
     5641 gaacataccg atgttttcac ggatttattg gaaacaaata gcagatgagg gcgtttttac
     5701 ttgtaatgaa ggcgaatgag aaggcaaggc ttacaggctt gttgtttatg ccaagcatca
     5761 atactcgcta tggacgagcg ccatcagcga taacaacaaa gtttgcaagt gcttgcttgg
     5821 atataaaagg cttgacgctt aagataaata gggccatact tgcttgcttt tactattttt
     5881 gttcctcata agcctgattg actacgctat tataaacccc tacattatcc tataatcaac
     5941 ttcacttgtg taacgttgaa tgagttcaac tgagtaatac tcggtcggaa atatacgctc
     6001 catctggcag gaaacatagg ctgcatttat atcttgcaca attatgccct tcgtaggtgt
     6061 atgttttcat gttttccatt atgtcaatgt agatgtatct ttggacaggc aacttttgga
     6121 tcaataaagc acaatttttg agcaatttga gatatccagt atttgtacgg gagtcccaag
     6181 attgtttagc ctaaaaggtt gaggacttgg attattcttc catcaagcgc ctccacccct
     6241 cgcatcctat ttcttttgaa gcatagggga cgagactgtc tgtaatttat ggcgaagtat
     6301 gacaagaaag attcaaactt tgttttacgt atgccccaat aaattctatg gtagcactac
     6361 tagagacgta ttttggtttt gtaataattt tcaaactgct cgctatgatt tgttacaaga
     6421 aacaaacaaa agacacgacc atagggttca tgttggtggt tgcgaagcca catctttaaa
     6481 acattactgc aaattcagca ggagtgagta gcaaaatcga caaaaactag agtacgcaac
     6541 attcccggtc gtaaatttct tatcgtggcg ccagaacatt cccgtccaat gatgagaatg
     6601 tctctaaaac aaatgcagtc acagagattt tctgcgactt ctcgagtgag aatgttgctt
     6661 attggtgcta gtggcttgag atcttcgtca agcaaattgg aatgccccca gattctcaaa
     6721 caagcacggc agaaactgaa atacggccaa aattatgaac ccttttgtgg gatctttgta
     6781 gaaataggac ctcaggatag tatgatgaag aatgttggca tgcaagcttg gcgtaatcat
     6841 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6901 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6961 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     7021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     7081 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     7141 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     7201 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7261 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7321 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7381 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7441 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7501 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7561 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7621 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7681 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7741 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7801 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     8041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     8101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     8161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     8221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8281 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8761 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8821 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8881 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8941 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctagctttt
     9001 tctttccaat tttttttttt tcgtcattat aaaaatcatt acgaccgaga ttcccgggta
     9061 ataactgata taattaaatt gaagctctaa tttgtgagtt tagctggccg catcttctca
     9121 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     9181 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     9241 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     9301 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     9361 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     9421 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     9481 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     9541 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     9601 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     9661 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     9721 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     9781 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     9841 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     9901 tttatcttcg tttcctgcag gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     9961 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
    10021 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
    10081 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagctc taagaaacca ttattatcat
    10141 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
