LOCUS       sequence-178549-        7840 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 7840)
  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 7840)
  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..7840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..44
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     261..283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     275..292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     276..292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     299..318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        299..317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1821..1837
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1821..1837
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      1958..2228
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     promoter        complement(2267..2285)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2306..2322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2306..2322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2319..2341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2330..2346
                     /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(2354..2360,2361..2378,2379..2384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2537..2554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2708..3296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3467..4258,4259..4327))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4090..4109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4328..4432)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4469..4972
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5014..5032
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5070..5092)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5192..5211
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(5704..6798)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(6811..7215)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(join(7516..7840,1..131))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7653..7674)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
       61 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
      121 acagggcgcg tcgcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      181 gcgggcctct tcgctattac gccagctggc gaagggggga tgtgctgcaa ggcgattaag
      241 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
      301 atacgactca ctatagggcg aattggagct ccaccgcggt ctaactgatc tatccaaaac
      361 tgaaaattac attcttgatt aggtttatca caggcaaatg taatttgtgg tattttgccg
      421 ttcaaaatct gtagaatttt ctcattggtc acattacaac ctgaaaatac tttatctaca
      481 atcataccat tcttataaca tgtcccctta atactaggat caggcatgaa cgcatcacag
      541 acaaaatctt cttgacaaac gtcacaattg atccctcccc atccgttatc acaatgacag
      601 gtgtcatttt gtgctcttat gggacgatcc ttattaccgc tttcatccgg tgatagaccg
      661 ccacagaggg gcagagagca atcatcacct gcaaaccctt ctatacactc acatctacca
      721 gtgtacgaat tgcattcaga aaactgtttg cattcaaaaa taggtagcat acaattaaaa
      781 catggcgggc acgtatcatt gcccttatct tgtgcagtta gacgcgaatt tttcgaagaa
      841 gtaccttcaa agaatggggt ctcatcttgt tttgcaagta ccactgagca ggataataat
      901 agaaatgata atatactata gtagagataa cgtcgatgac ttcccatact gtaattgctt
      961 ttagttgtgt atttttagtg tgcaagtttc tgtaaatcga ttaatttttt tttctttcct
     1021 ctttttatta accttaattt ttattttaga ttcctgactt caactcaaga cgcacagata
     1081 ttataacatc tgcacaatag gcatttgcaa gaattactcg tgagtaagga aagagtgagg
     1141 aactatcgca tacctgcatt taaagatgcc gatttgggcg cgaatccttt attttggctt
     1201 caccctcata ctattatcag ggccagaaaa aggaagtgtt tccctccttc ttgaattgat
     1261 gttaccctca taaagcacgt ggcctcttat cgagaaagaa attaccgtcg ctcgtgattt
     1321 gtttgcaaaa agaacaaaac tgaaaaaacc cagacacgct cgacttcctg tcttcctatt
     1381 gattgcagct tccaatttcg tcacacaaca aggtcctagc gacggctcac aggttttgta
     1441 acaagcaatc gaaggttctg gaatggcggg aaagggttta gtaccacatg ctatgatgcc
     1501 cactgtgatc tccagagcaa agttcgttcg atcgtactgt tactctctct ctttcaaaca
     1561 gaattgtccg aatcgtgtga caacaacagc ctgttctcac acactctttt cttctaacca
     1621 agggggtggt ttagtttagt agaacctcgt gaaacttaca tttacatata tataaacttg
     1681 cataaattgg tcaatgcaag aaatacatat ttggtctttt ctaattcgta gtttttcaag
     1741 ttcttagatg ctttcttttt ctctttttta cagatcatca aggaagtaat tatctacttt
     1801 ttacaacaaa tataaaacaa tctagaacta gtggatcccc cgggctgcag gaattcgata
     1861 tcaagcttcc catgtctcta ctggtggtgg tgcttctttg gaattattgg aaggtaagga
     1921 attgccaggt gttgctttct tatccgaaaa gaaataaatt gaattgaatt gaaatcgata
     1981 gatcaatttt tttcttttct ctttccccat cctttacgct aaaataatag tttattttat
     2041 tttttgaata ttttttattt atatacgtat atatagacta ttatttatct tttaatgatt
     2101 attaagattt ttattaaaaa aaaattcgct cctcttttaa tgcctttatg cagttttttt
     2161 ttcccattcg atatttctat gttcgggttc agcgtatttt aagtttaata actcgaaaat
     2221 tctgcgttcg ttactcgagg gggggcccgg tacccagctt ttgttccctt tagtgagggt
     2281 taattccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2341 tcacaattcc acacaacata ggagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2401 gagtgaggta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2461 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2521 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2581 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2641 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2701 tggcgttttt ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc
     2761 agaggtggcg aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc
     2821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2881 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     2941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3121 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     3181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3241 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactgc
     3541 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3601 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3661 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3721 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3781 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3841 aacgatcaag gcgagttaca tgatccccca tgttgtgaaa aaaagcggtt agctccttcg
     3901 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3961 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4021 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4081 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4141 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4201 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4261 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4321 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4441 cccgaaaagt gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa
     4501 ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa
     4561 tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt
     4621 ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag
     4681 aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat
     4741 ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa
     4801 ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt
     4861 tactttctaa aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa
     4921 ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtcctaa
     4981 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5041 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5101 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5161 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5221 caccatatcg actacgtcgt aaggccgttt ctgacagagt aaaattcttg agggaacttt
     5281 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
     5341 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatcaaa
     5401 ttaggaatcg tagtttcatg attttctgtt acacctaact ttttgtgtgg tgccctcctc
     5461 cttgtcaata ttaatgttaa agtgcaattc tttttcctta tcacgttgag ccattagtat
     5521 caatttgctt acctgtattc ctttactatc ctcctttttc tccttcttga taaatgtatg
     5581 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
     5641 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
     5701 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
     5761 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
     5821 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
     5881 caagatagtg gcgatagggt caaccttatt ctttggcaaa tctggagcag aaccgtggca
     5941 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
     6001 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     6061 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     6121 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
     6181 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     6241 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     6301 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     6361 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     6421 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     6481 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     6541 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat caaccttctt
     6601 ggaggcttcc agcgcctcat ctggaagtgg gacacctgta gcatcgatag cagcaccacc
     6661 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
     6721 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
     6781 cttcttaggg gcagacatag gggcagacat tagaatggta tatccttgaa atatatatat
     6841 atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta
     6901 ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca
     6961 tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcctc tcacctttcc
     7021 tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac
     7081 aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta
     7141 tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg
     7201 agtattccca cagttaactg cggtcaagat atttcttgaa tcaggcgcct tagaccgctc
     7261 ggccaaacaa ccaattactt gttgagaaat agagtataat tatcctataa atataacgtt
     7321 tttgaacaca catgaacaag gaagtacagg acaattgatt ttgaagagaa tgtggatttt
     7381 gatgtaattg ttccgattcc atttttaata aggcaataat attaggtatg tggatatact
     7441 agaagttctc ctcgaccggt cgatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7501 aaataccgca tcaggaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     7561 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7621 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7681 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7741 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7801 accctaaagg gagcccccga tttagagctt gacggggaaa
//
