LOCUS       sequence-202537-        6773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for pop-in/pop-out replacement of Snx4 with Snx4-iGFPx3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6773)
  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 6773)
  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-SNX4-iGFPx3
FEATURES             Location/Qualifiers
     source          1..6773
                     /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(3271..3288)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3271..3287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3280..3302)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3496..3515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3615..3637
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3755..4558)
                     /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(4559..4774)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4781..4799)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4867..4971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4972..5040,5041..5832)
                     /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(5190..5209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6003..6591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6492..6511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6745..6762
                     /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 atgcgaaaac gaggaacttg ctgcgaagtt gaagattttt aatgaaggtg taacacagct
      301 gtcatatggc ttgcaagatc ttacaaagta tctggattac gaatatattg ttgatttgaa
      361 ggatcttgaa cactacattg atagtatgcg acagctgatc aaattgaaag accagaaaca
      421 gatagactac gaagaactaa gtgattattt aaccaggtca atcaaagaaa aaaataactt
      481 gatttctgga tacggaggta gtaatttctt tgccaataaa ttggaagaac tagcgggaat
      541 aaatcaggag gcttcacgta gagaaaaaat caataaacta gaaggcaaaa taacgtcatt
      601 aacaggtgaa ttagaaaacg ctaaaaaggt ggcggatgga ttcgaacagg aatgcttgaa
      661 ggaaatagac catttcgaaa gtgtaaaaac ggctgaaatc aagaaatctc tgggctcact
      721 agcggaccat catatcgaat tctacgaaag gatacttgaa gcatgggaaa aagtagacga
      781 tagtctaacg gatccaggaa gttcggggtc cagcggttct tcaggcgcta gcatggatag
      841 cactgagagc cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
      901 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
      961 ccttaaattt atttgcacta ctggaaaact acctgttcct tggccaacac ttgtcactac
     1021 tttaacttat ggtgttcagt gcttttcaag atacccggat catatgaaac agcatgactt
     1081 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     1141 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     1201 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttgggcaca aattggaata
     1261 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagc
     1321 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     1381 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     1441 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttaaagagtt
     1501 tgtaacagct gctgggatta cacatggcag ctccggctcc agcgggtcta gcatggatag
     1561 cactgagagc cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     1621 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     1681 ccttaaattt atttgcacta ctggaaaact acctgttcct tggccaacac ttgtcactac
     1741 tttaacttat ggtgttcagt gcttttcaag atacccggat catatgaaac agcatgactt
     1801 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     1861 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     1921 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttgggcaca aattggaata
     1981 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagc
     2041 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     2101 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2161 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttaaagagtt
     2221 tgtaacagct gctgggatta cacatggcag ctccggctcc agcgggtcta gcatggatag
     2281 cactgagagc cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     2341 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     2401 ccttaaattt atttgcacta ctggaaaact acctgttcct tggccaacac ttgtcactac
     2461 tttaacttat ggtgttcagt gcttttcaag atacccggat catatgaaac agcatgactt
     2521 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     2581 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     2641 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttgggcaca aattggaata
     2701 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagc
     2761 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     2821 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     2881 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttaaagagtt
     2941 tgtaacagct gctgggatta cacatggcag ctccggctcc agcgggtaaa gcggccgctg
     3001 gtgctcttaa tgttttccca ttactactga taataccttg ggcactacga ttatctgatc
     3061 acatttaaca gagaaattta aaaaaagaaa aatatctata taaaaatatg taaatgttgc
     3121 tattatttta tgatgttctg atttttttcg tcaaacctta gatttcttat gtacattaaa
     3181 cttaaacatt ttgcattgtt cttccagttc ttggattata ttatcttggt gggagtcaat
     3241 caaatcccaa tcggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     3301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3361 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3421 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3481 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3541 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3601 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3661 cgcgagacga aagggcgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     3721 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     3781 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     3841 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     3901 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     3961 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     4021 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     4081 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     4141 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     4201 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     4261 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     4321 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     4381 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     4441 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     4501 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     4561 tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     4621 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     4681 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     4741 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagctg gcctcgtgat acgcctattt
     4801 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     4861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aga
//
