LOCUS       sequence-202389-        5676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Arf1 with Arf1-msGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5676)
  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 5676)
  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-ARF1-msGFP
FEATURES             Location/Qualifiers
     source          1..5676
                     /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(1168..1189)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1776..1797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2174..2191)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2174..2190)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2183..2205)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2399..2418)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2518..2540
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2658..3461)
                     /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(3462..3677)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3684..3702)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3770..3874
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3875..3943,3944..4735)
                     /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(4093..4112)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4906..5494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5395..5414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5648..5665
                     /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 ccaagcttga
      241 tagcagttta ttgtagaaaa accatgttat tacccttccc tttttatttc ttttcgcgtt
      301 gcaaatcaca tataacgagg tggcttgtat ttgtcaaacc aaaaaaaaaa atgaaaatcg
      361 aaaaatggaa aaacagagag agaaacggaa tctttgacac gttctcgatc cacttgttta
      421 tcgaggtggt ttttataagt cttactagat agaaagttca ttttgttttg aaactttttg
      481 gaacctctgg cattgaaggt ataagaaaga actcaaacag gtttaataga attaaaaatg
      541 ggtttgtttg cctctaagtt gttcagtaac ctttttggta acaaagaaat gcgtattctt
      601 atggttggtc ttgatggtgc tggtaagacc accgttttgt acaagttgaa attgggtgaa
      661 gttatcacta ccattccaac aattggtttc aacgttgaaa ctgtccaata taagaacatt
      721 tcattcactg tctgggatgt cggtggacaa gacagaatta gatctctatg gagacactac
      781 tacagaaaca ctgaaggtgt tatctttgtt gtcgattcta acgatagatc gcgtattggt
      841 gaagctagag aagttatgca aagaatgttg aacgaagatg aattgagaaa cgccgcttgg
      901 ttggtgttcg ctaacaagca agatttgcca gaagccatgt ctgctgctga aatcactgaa
      961 aaactaggtt tacattctat tagaaaccgt ccatggttta tccaagccac gtgtgctacc
     1021 tccggtgaag gtttgtatga aggtttggaa tggttaagta acagtttgaa aaactcaact
     1081 acggatccag gaagttcggg gtccagcggt tcttcaggcg ctagcatgga tagcactgag
     1141 agcctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1201 aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
     1261 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     1321 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     1381 tccgccatgc ccgaaggcta cgtccaggag cgcaccatca ccttcaagga cgacggcacc
     1441 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1501 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
     1561 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     1621 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1681 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1741 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1801 gccgccggga tcactcacgg cagctccggc tccagcgggt aaagcggccg caaattctag
     1861 aatatggatc aaatacgctt gtataaacta aatgaaacat aaagattaag aacttaagag
     1921 gccaacgtcg atggatttat tgacgatcac cagccaacac atatagattt tagtgtaaaa
     1981 gcaataaaaa ccaagataat aaaataaaaa aatactgaag aagcctaact agtataaact
     2041 actttaacta ataatggcaa tttgatatag aaacaaagaa acatgatata tttaggatat
     2101 tatacaacgc attttcattt gttttacagc acccctgcgt gaatcatata ttgacggtac
     2161 cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2221 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2281 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2341 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     2401 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2461 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2521 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcg
     2581 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     2641 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     2701 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     2761 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     2821 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     2881 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     2941 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     3001 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     3061 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     3121 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     3181 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     3241 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     3301 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     3361 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     3421 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     3481 catgtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     3541 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     3601 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     3661 aaaaatgatg aattgaaaag ctggcctcgt gatacgccta tttttatagg ttaatgtcat
     3721 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3781 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3841 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3901 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     3961 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4021 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4081 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4141 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4201 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4261 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4321 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4381 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4441 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4501 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4561 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4621 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4681 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4741 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4801 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5041 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5521 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5581 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5641 cgagcgcagc gagtcagtga gcgaggaagc ggaaga
//
