LOCUS       sequence-138754-        8894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Crepis alpina delta-12 fatty acid acetylenase (vFAD2) with 
            C-terminal FLAG epitope and delta-15 fatty acid desaturase (FAD3) 
            with C-terminal c-myc epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8894)
  AUTHORS   Kappock
  TITLE     Additional fatty acid desaturase and acetylenase constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8894)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pJK372
FEATURES             Location/Qualifiers
     source          1..8894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      69..1411
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    401..448
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1438..1542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1543..1611,1612..2403)
                     /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(1761..1780)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3063..3082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3316..3333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3405..3594)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(3624..3653)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4676..4687
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        4830..5494
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(4843..4866)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5161..5278
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             6674..6697
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6845..7010
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6900..6924)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      7089..7544
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7176..7195)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7386..7407
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7675..8478)
                     /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(8479..8699)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8725..8744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8844..8866
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggatcttcga gcgtcccaaa
     3421 accttctcaa gcaaggtttt cagtataatg ttacatgcgt acacgcgtct gtacagaaaa
     3481 aaaagaaaaa tttgaaatat aaataacgtt cttaatacta acataactat aaaaaaataa
     3541 atagggacct agacttcagg ttgtctaact ccttcctttt cggttagagc ggatcttagc
     3601 tagccgcggt accaagctta ctcgaggtct tcttcggaaa tcaacttctg ttccatgtcg
     3661 acctcaattt tcttcccaat cattcgataa tcagtttgat agtaaacgac ttcacccgca
     3721 tcactaacat aatgatcttg tttgacgctt ttcaccaagt tatccattaa gtgaaccggt
     3781 atccagccag acttttttgg ctctcggtag taattcccaa gcacagactt tgccgccatt
     3841 gtcgcttcaa tcagatgata atgtgggatt tgtgggaaaa gatgatggat gacatgggtg
     3901 cctatgtcat gatggatgtt attgaatatt ccataatctc gatcaacggt tgttaatcct
     3961 ccccttaggt agctccattc tttgccacga taccaaggaa gtttttgctc atggccgtgg
     4021 tgatgcaaat aagtgacaaa atccaaccac ataacgaata tcaaataagg aatgccatag
     4081 aggttgaaca atagagttgg gccaataaca attgacaaaa ataccagatt ggctaccatt
     4141 agagaccaac ataaagtaga tgtcacaata taacttcttt cattggggga gaacatatca
     4201 ctatacgggt tgaaatgtga ccctgatttg cctgggctcc tataccacaa gtacaatggg
     4261 taggcaaata agggaaaagg gactttgaat ctgagtagtt tggtggaagc atccaatgtg
     4321 ttgtaagtct tttcggtcaa cggaacccat gattcatcat tttcgacatg accatgattt
     4381 tgatgatggg tcctgtggct gattctccac ccatggtagg gtacgagaat cgaagaatgc
     4441 aaaatgtggc ccacgacgct attgagatta gggttgtctg aaaagcttcc atggccacaa
     4501 tcatgaccaa gaacaaagat tgcccagaac attgtgcctt gtgcaatcca gtaaatcggc
     4561 caaacccatg aactggtttt gaagataaca gcgatggcga ccaatgagga aattaccaga
     4621 atgtccctga ggacgtagct gagagacctc catggattct tgacccagca atggctggga
     4681 attgcagctc tgatgtcagc caccttgaat ggaggagggg cacttgggtc gaattcttct
     4741 tgattctttt ggacgccatt gacgagaggc gaagcttgat taagcttttc tgtggcatct
     4801 ggagaagaaa ccgccattgt ggatccgggg ttttttctcc ttgacgttaa agtatagagg
     4861 tatattaaca attttttgtt gatactttta ttacatttga ataagaagta atacaaaccg
     4921 aaaatgttga aagtattagt taaagtggtt atgcagtttt tgcatttata tatctgttaa
     4981 tagatcaaaa atcatcgctt cgctgattaa ttaccccaga aataaggcta aaaaactaat
     5041 cgcattatca tcctatggtt gttaatttga ttcgttcatt tgaaggtttg tggggccagg
     5101 ttactgccaa tttttcctct tcataaccat aaaagctagt attgtagaat ctttattgtt
     5161 cggagcagtg cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg aagacgagga
     5221 cgcacggagg agagtcttcc ttcggagggc tgtcacccgc tcggcggctt ctaatccgta
     5281 cttcaatata gcaatgagca gttaagcgta ttactgaaag ttccaaagag aaggtttttt
     5341 taggctaaga taatggggct ctttacattt ccacaacata taagtaagat tagatatgga
     5401 tatgtatatg gatatgtata tggtggtaat gccatgtaat atgattatta aacttctttg
     5461 cgtccatcca aaaaaaaagt aagaattttt gaaaattcga attcaaccct cactaaaggg
     5521 cggccgcact agtatgggtg gcggtggccg tggtcggact tcgcaaaaac ccctcatgga
     5581 acgtgtctca gttgatccac ccttcaccgt gagtgatctc aagcaagcaa tccctcccca
     5641 ttgcttcaag cgatctgtaa tccgttcctc ttactacata gtccacgatg ctattatcgc
     5701 ctacatcttc tacttccttg ccgacaaata cattccgatt ctccctgccc ctctagccta
     5761 cctcgcttgg cccctttact ggttctgtca agctagcatc ctcaccggct tatgggtcat
     5821 cggtcacgaa tgcggtcacc atgccttcag cgactaccag tgggttgacg acactgtggg
     5881 cttcatcctc cactcgtttc tcatgacccc gtatttctcc tggaaataca gccaccggaa
     5941 ccaccatgcc aacacaaatt cgcttgacaa cgatgaagtt tacatcccca aaagcaaggc
     6001 caaagtcgcg ctttactata aagttctcaa ccacccacct ggccgactgt tgattatgtt
     6061 catcaccttc accctaggct tccctctata cctctttacc aatatttccg gcaagaagta
     6121 tgaaaggttt gccaaccatt tcgaccccat gagtccgatt ttcaaagagc gtgagcggtt
     6181 tcaggtcttg ctatcggatc ttggccttct tgctgtgctt tacggagtta aacttgcggt
     6241 agcagcgaaa ggcgccgctt gggtgacgtg catttacgga attccagttt taggcgtgtt
     6301 tatctttttc gatatcatca cctacttgca ccacacccat ctgtcgttgc ctcattatga
     6361 ttcatctgaa tggaactggc tcagaggggc tttgtcaaca atcgataggg actttgggtt
     6421 cctgaatagt gtgctccatg atgttacaca cactcacgtt atgcatcatc tgttttcata
     6481 cattccacac tatcatgcga aggaggcaag ggatgcaatc aacacagtct tgggcgactt
     6541 ttataagatc gataggactc caattctgaa agcaatgtgg agagaggcca aggaatgcat
     6601 cttcatcgag cctgaaaaag gtagggagtc caagggtgta tattggtaca ataaattcgc
     6661 actagtatcg atggattaca aggatgacga cgataagatc tgagctctta attaacaatt
     6721 cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta ccttgccaga
     6781 aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg acacttctaa
     6841 ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa aaaaaataag
     6901 tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat tcttgagtaa
     6961 ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc caattcagct
     7021 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     7081 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     7141 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     7201 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     7261 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     7321 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     7381 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     7441 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     7501 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     7561 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
     7621 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     7681 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     7741 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     7801 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     7861 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     7921 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     7981 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     8041 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     8101 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     8161 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     8221 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     8281 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     8341 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     8401 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     8461 atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     8521 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     8581 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     8641 atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaattgaaa
     8701 agctgtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     8761 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     8821 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     8881 accgaaacgc gcga
//
