LOCUS       sequence-202396-        5958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating construct for expressing GFP-Snc1(L96V) from the TPI1 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5958)
  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 5958)
  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: pRS406-GFP-SNC1(L96V)
FEATURES             Location/Qualifiers
     source          1..5958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(144..149,150..167,168..174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..198
                     /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     187..209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     206..222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     206..222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     241..261
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        243..261
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        306..717
                     /gene="S. cerevisiae TPI1"
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose 
                     phosphate isomerase"
     CDS             728..1441
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(758..786)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1381..1401
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2003..2020)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2003..2019)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2012..2034)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2160..2615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2247..2266)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2457..2478
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2746..3549)
                     /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(3550..3765)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3789..3808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3908..3930
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3968..3986)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4054..4158
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4159..4227,4228..5019)
                     /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(4377..4396)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5190..5778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5679..5698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5932..5949
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
      241 gcaattaacc ctcactaaag ggaacaaaag ctggagctcc accgcggtgg cggccccccc
      301 tcgagccatc aggttggtgg aagattaccc gttctaagac ttttcagctt cctctattga
      361 tgttacacct ggacacccct tttctggcat ccagttttta atcttcagtg gcatgtgaga
      421 ttctccgaaa ttaattaaag caatcacaca attctctcgg ataccacctc ggttgaaact
      481 gacaggtggt ttgttacgca tgctaatgca aaggagccta tatacctttg gctcggctgc
      541 tgtaacaggg aatataaagg gcagcataat ttaggagttt agtgaacttg caacatttac
      601 tattttccct tcttacgtaa atatttttct ttttaattct aaatcaatct ttttcaattt
      661 tttgtttgta ttcttttctt gcttaaatct ataactacaa aaaacacata cataaacaag
      721 ctagcttatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      781 agatggtgat gttaatggac acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      841 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      901 aacacttgtc actactttct cttatggtgt tcaatgcttt tcaagatacc cagatcatat
      961 gaagcggcac gacttcttca agagcgccat gcctgaggga tacgtgcagg agaggaccat
     1021 cttcttcaag gacgacggga actacaagac acgtgctgaa gtcaagtttg agggagacac
     1081 cctcgtcaac aggatcgagc ttaagggaat cgatttcaag gaggacggaa acatcctcgg
     1141 ccacaagttg gaatacaact acaactccca caacgtatac atcatggccg acaaacaaaa
     1201 gaatggaatc aaagctaact tcaaaactag acacaacatt gaagatggag gcgttcaact
     1261 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     1321 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     1381 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     1441 atcgaattcc atgtcgtcat ctactccctt tgacccttat gctctatccg agcacgatga
     1501 agaacgaccc cagaatgtac agtctaagtc aaggactgcg gaactgcagg ctgaaattga
     1561 tgataccgtg ggaataatga gagataacat aaataaagta gctgagcgag gtgaaagatt
     1621 aacgtccatt gaagataaag ccgataacct agcggtctca gcccaaggct ttaagagggg
     1681 tgccaatagg gtcagaaaag ccatgtggta caaggatcta aaaatgaaga tgtgtgtggc
     1741 tttagtaatc atcatattgc ttgttgtaat catcgtcccc attgctgttc actttagtcg
     1801 atagaagttc actcgcaatg ctatatatat agggagcttc catatgtagt gtaggatata
     1861 tgtatatata catatataca tatatgggcg tatatttact ttgttcttat tttctgtcta
     1921 attttataat tttactgaca gtagctaagc cctctgtatt gctgggatcc ccatggattc
     1981 aaagccgata ccaaacgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     2041 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     2101 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga
     2161 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2221 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2281 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2341 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2401 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2461 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2521 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2581 cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg
     2641 catctgtgcg gtatttcaca ccgcataggg taataactga tataattaaa ttgaagctct
     2701 aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt tttgctggcc
     2761 gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct ctaccttagc
     2821 atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc ctgtagagac
     2881 cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat ctaaacccac
     2941 accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt ctctttgagc
     3001 aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac ccttagtata
     3061 ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa ggcctctagg
     3121 ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg ggcccaccac
     3181 accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg cagagtactg
     3241 caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta
     3301 cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat cagtcaagat
     3361 atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta actccagtaa
     3421 ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt cgtgcatgat
     3481 attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct tatatgtagc
     3541 tttcgacatg atttatcttc gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa
     3601 tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca atctaagtct
     3661 gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa aatttcaagg
     3721 aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaaggt ggtatggtgc
     3781 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     3841 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     3901 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     3961 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4021 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4081 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4141 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4201 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4261 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4321 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4381 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4441 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4501 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4561 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4621 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4681 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4741 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4801 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4861 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4921 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4981 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5041 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5101 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5161 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5221 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5281 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5341 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5401 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5461 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5521 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5581 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5641 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5701 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5761 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5821 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5881 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5941 gtgagcgagg aagcggaa
//
