LOCUS       sequence-189323-        8928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GFP-tagged Arabidopsis SEIPIN1 gene in yeast cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8928)
  AUTHORS   Cai Y, Goodman JM, Pyc M, Mullen RT, Dyer JM, Chapman KD
  TITLE     Arabidopsis SEIPIN Proteins Modulate Triacylglycerol Accumulation 
            and Influence Lipid Droplet Proliferation.
  JOURNAL   Plant Cell. 2015 Sep;27(9):2616-36. doi: 10.1105/tpc.15.00588. Epub 
            2015 Sep 11.
  PUBMED    26362606
REFERENCE   2  (bases 1 to 8928)
  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: 
            "10.1105/tpc.15.00588"; journalName: "Plant Cell"; date: "2015-09"; 
            volume: "27"; issue: "9"; pages: "2616-36"
COMMENT     Sequence Label: pRS315-PGK-SEIPIN1
FEATURES             Location/Qualifiers
     source          1..8928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(54..71)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(225..813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(305..324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(984..1775,1776..1844))
                     /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     1607..1626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1845..1949)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1986..2489
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2531..2549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2587..2609)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2709..2728
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3222..4316)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4317..4724)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(5025..5480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5162..5183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5374..5393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5606..5628
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5620..5637
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5621..5637
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5644..5663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5644..5662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7166..7182
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7166..7182
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      8404..8674
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     promoter        complement(8712..8730)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8751..8767)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8751..8767)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8764..8786)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8775..8791
                     /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(8799..8805,8806..8823,8824..8829))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8844..8865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
       61 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
      121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
      181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
      241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
      301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
      361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
      421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
      481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
      541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
      601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
      661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
      721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
      781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
      841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     1081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     1141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     1201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     1261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     1321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     1381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     1441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     1501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     1561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     1621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     1681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     1741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc
     1981 ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa atatataaat
     2041 taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt
     2101 aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca
     2161 ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt
     2221 gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc
     2281 taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt
     2341 tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa
     2401 aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg
     2461 aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa accattatta tcatgacatt
     2521 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     2581 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2641 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     2701 taactatgcg gcatcagagc agattgtact gagagtgcac catatcgact acgtcgttaa
     2761 ggccgtttct gacagagtaa aattcttgag ggaactttca ccattatggg aaatggttca
     2821 agaaggtatt gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt ttatttgttg
     2881 tatttttttt tttttagaga aaatcctcca atatataaat taggaatcat agtttcatga
     2941 ttttctgtta cacctaactt tttgtgtggt gccctcctcc ttgtcaatat taatgttaaa
     3001 gtgcaattct ttttccttat cacgttgagc cattagtatc aatttgctta cctgtattcc
     3061 tttacatcct cctttttctc cttcttgata aatgtatgta gattgcgtat atagtttcgt
     3121 ctaccctatg aacatattcc attttgtaat ttcgtgtcgt ttctattatg aatttcattt
     3181 ataaagttta tgtacaaata tcataaaaaa agagaatctt tttaagcaag gattttctta
     3241 acttcttcgg cgacagcatc accgacttcg gtggtactgt tggaaccacc taaatcacca
     3301 gttctgatac ctgcatccaa aaccttttta actgcatctt caatggcctt accttcttca
     3361 ggcaagttca atgacaattt caacatcatt gcagcagaca agatagtggc gatagggttg
     3421 accttattct ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa accaaatgcg
     3481 gtgttcttgt ctggcaaaga ggccaaggac gcagatggca acaaacccaa ggaacctggg
     3541 ataacggagg cttcatcgga gatgatatca ccaaacatgt tgctggtgat tataatacca
     3601 tttaggtggg ttgggttctt aactaggatc atggcggcag aatcaatcaa ttgatgttga
     3661 accttcaatg tagggaattc gttcttgatg gtttcctcca cagtttttct ccataatctt
     3721 gaagaggcca aaacattagc tttatccaag gaccaaatag gcaatggtgg ctcatgttgt
     3781 agggccatga aagcggccat tcttgtgatt ctttgcactt ctggaacggt gtattgttca
     3841 ctatcccaag cgacaccatc accatcgtct tcctttctct taccaaagta aatacctccc
     3901 actaattctc tgacaacaac gaagtcagta cctttagcaa attgtggctt gattggagat
     3961 aagtctaaaa gagagtcgga tgcaaagtta catggtctta agttggcgta caattgaagt
     4021 tctttacgga tttttagtaa accttgttca ggtctaacac taccggtacc ccatttagga
     4081 ccacccacag cacctaacaa aacggcatca gccttcttgg aggcttccag cgcctcatct
     4141 ggaagtggaa cacctgtagc atcgatagca gcaccaccaa ttaaatgatt ttcgaaatcg
     4201 aacttgacat tggaacgaac atcagaaata gctttaagaa ccttaatggc ttcggctgtg
     4261 atttcttgac caacgtggtc acctggcaaa acgacgatct tcttaggggc agacattaga
     4321 atggtatatc cttgaaatat atatatatat attgctgaaa tgtaaaaggt aagaaaagtt
     4381 agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt aaataatatt
     4441 gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta ttctatatga
     4501 aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt tgaaaaaggt
     4561 atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg
     4621 tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag
     4681 attcgaactc ttgcatctta cgatacctga gtattcccac agttaactgc ggtcaagata
     4741 tttcttgaat caggcgcctt agaccgctcg gccaaacaac caattacttg ttgagaaata
     4801 gagtataatt atcctataaa tataacgttt ttgaacacac atgaacaagg aagtacagga
     4861 caattgattt tgaagagaat gtggattttg atgtaattgt tgggattcca tttttaataa
     4921 ggcaataata ttaggtatgt agatatacta gaagttctcc tcgaccggtc gatatgcggt
     4981 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa
     5041 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5101 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5161 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5221 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5281 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5341 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5401 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5461 tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5521 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5581 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     5641 ttgtaatacg actcactata gggcgaattg gagctccacc gcggtctaac tgatctatcc
     5701 aaaactgaaa attacattct tgattaggtt tatcacaggc aaatgtaatt tgtggtattt
     5761 tgccgttcaa aatctgtaga attttctcat tggtcacatt acaacctgaa aatactttat
     5821 ctacaatcat accattctta taacatgtcc ccttaatact aggatcaggc atgaacgcat
     5881 cacagacaaa atcttcttga caaacgtcac aattgatccc tccccatccg ttatcacaat
     5941 gacaggtgtc attttgtgct cttatgggac gatccttatt accgctttca tccggtgata
     6001 gaccgccaca gaggggcaga gagcaatcat cacctgcaaa cccttctata cactcacatc
     6061 taccagtgta cgaattgcat tcagaaaact gtttgcattc aaaaataggt agcatacaat
     6121 taaaacatgg cgggcatgta tcattgccct tatcttgtgc agttagacgc gaatttttcg
     6181 aagaagtacc ttcaaagaat ggggtcttat cttgttttgc aagtaccact gagcaggata
     6241 ataatagaaa tgataatata ctatagtaga gataacgtcg atgacttccc atactgtaat
     6301 tgcttttagt tgtgtatttt tagtgtgcaa gtttctgtaa atcgattaat ttttttttct
     6361 ttcctctttt tattaacctt aatttttatt ttagattcct gacttcaact caagacgcac
     6421 agatattata acatctgcat aataggcatt tgcaagaatt actcgtgagt aaggaaagag
     6481 tgaggaacta tcgcatacct gcatttaaag atgccgattt gggcgcgaat cctttatttt
     6541 ggcttcaccc tcatactatt atcagggcca gaaaaaggaa gtgtttccct ccttcttgaa
     6601 ttgatgttac cctcataaag cacgtggcct cttatcgaga aagaaattac cgtcgctcgt
     6661 gatttgtttg caaaaagaac aaaactgaaa aaacccagac acgctcgact tcctgtcttc
     6721 ctattgattg cagcttccaa tttcgtcaca caacaaggtc ctagcgacgg ctcacaggtt
     6781 ttgtaacaag caatcgaagg ttctggaatg gcgggaaagg gtttagtacc acatgctatg
     6841 atgcccactg tgatctccag agcaaagttc gttcgatcgt actgttactc tctctctttc
     6901 aaacagaatt gtccgaatcg tgtgacaaca acagcctgtt ctcacacact cttttcttct
     6961 aaccaagggg gtggtttagt ttagtagaac ctcgtgaaac ttacatttac atatatataa
     7021 acttgcataa attggtcaat gcaagaaata catatttggt cttttctaat tcgtagtttt
     7081 tcaagttctt agatgctttc tttttctctt ttttacagat catcaaggaa gtaattatct
     7141 actttttaca acaaatataa aacaatctag aactagtgga tccatgagaa tcctccaaaa
     7201 caaaacaatg aaagaacaag acaaccaact caaaatccct gaaccattac gagctgactg
     7261 gttcatggtt ctagtcacca tccaagctga tctaatctac aacgcactcg tggtattatc
     7321 ctcacctttc tttctcctat accgctctta ccgccgcgca gtcgttaccg tctccgcagc
     7381 agaaaaagcg gttaaacgtg ctccagcgca gatagctggt ggtgcggggc gggttgttag
     7441 gaggacatgg tttggtatat tgggtgcatg ccacgtgtca atggtaatgg tgttggcttt
     7501 gattttggcg gttgttattg gtgttgggat cgtgagtttg tatgtagaaa agcctgtggt
     7561 tgtgagagac agattgttct ttgattacac tgaagaaaac cccagtgcgg tttttagttt
     7621 tgataagaag aagagatcgt ttagtgttcc tgttggtcat agtgttcatg tctctttagt
     7681 gctttggatg cctgaatctg aaattaatcg aagaattgga gtatttcagt tgaaagtaga
     7741 gcttttatcg ttgaaaggcg agacaatcgc gagatcgagc caaccttgta tgttacggtt
     7801 cagaagcaaa cctataagac ttgcacgaac atttgtgatg agtgttccac ttatagccgg
     7861 gatagcaaac gaagcacaaa caatgagaat cgatgcatta aagcaccaag agaaaatgcc
     7921 aaggacaaaa gccgtgagag cgactctgat tccgcgtgca caaactcgga cattgcctca
     7981 gttatatgaa gcagagatcg ttataaactc aaaaccgccc tggattaaac gaatggctta
     8041 taactggaaa tggactcttt gcgtatggac ttcaatgtac ctttacgtcg cgattttgac
     8101 cgcacttctt tggtgtttta gaccggtttt gttcccttac acttcctcaa gaaccatttc
     8161 ggagagcgaa aacttagaga tcgaagtagt tgaagaagaa caagaacaag tcatggaaag
     8221 acgacggagg gaaagaagaa accagcctcg tcgaagaaat tttgctacta cacagaaaag
     8281 ttatacatag ctgcaggaat tcgatatcaa gcttcccatg tctctactgg tggtggtgct
     8341 tctttggaat tattggaagg taaggaattg ccaggtgttg ctttcttatc cgaaaagaaa
     8401 taaattgaat tgaattgaaa tcgatagatc aatttttttc ttttctcttt ccccatcctt
     8461 tacgctaaaa taatagttta ttttattttt tgaatatttt ttatttatat acgtatatat
     8521 agactattat ttatctttta atgattatta agatttttat taaaaaaaaa ttcgctcctc
     8581 ttttaatgcc tttatgcagt ttttttttcc cattcgatat ttctatgtac gggttcagcg
     8641 tattttaagt ttaataactc gaaaattctg cgttcgttac tcgagggggg gcccggtacc
     8701 agcttttgtt ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg
     8761 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     8821 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     8881 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatga
//
