LOCUS       sequence-139406-        8870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Crepis alpina delta-12 fatty acid acetylenase (vFAD2) with 
            C-terminal FLAG epitope and delta-12 fatty acid desaturase (FAD2-3) 
            with C-terminal c-myc epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8870)
  AUTHORS   Kappock
  TITLE     Additional fatty acid desaturase and acetylenase constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8870)
  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            "Unpublished"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: pJK371
FEATURES             Location/Qualifiers
     source          1..8870
                     /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"
     promoter        4806..5470
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(4819..4842)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5137..5254
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             6650..6673
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6821..6986
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6876..6900)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      7065..7520
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7152..7171)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7362..7383
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7651..8454)
                     /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(8455..8675)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8701..8720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8820..8842
                     /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 acatatttat tcatgtacca aaagacaccc cctttcttct catccgactc cacatacaaa
     3721 cactctttcg cctccctcca cattgccaca taaaacggcg tatcatcaag ccgataatac
     3781 tccccaagaa ccggcttcac tgctttcgtt gcttccatgg cataataatg aggcattgta
     3841 gagaacaaat ggtgcaccac atgcgtatca gtgatattat gaaacacctt attcaacaca
     3901 ccatagtccc gatccacggt tgccaaagcc ccccttaacc aatcccactc cgaatcatca
     3961 tagtgaggca gcgaagggtg cgtatgttga aggtatgtaa tcataaccag aaacccgttt
     4021 acaataagca acgggacacc gtacacggaa atgagccaag ttagaccttt gagcatggcg
     4081 atacggtaaa gtagatatga aattatgata ataccaccat cggatatcca aatttggaga
     4141 cgttcgcggg tgttgaaaat agggctgttt ggagcgaagt ggttagcaaa acgatcgtat
     4201 ggtctacctg agacgttgaa tgctaggtac aatggccgac caagaataag agtggtgaga
     4261 agagttaata aacgtccggg agggtagttt aggtattttg cgtagcatga taatttcgat
     4321 ttaggtttag ggacaaaaac ctcgtcacgc tcaagtgagg cggtgtcgga gtggtggcga
     4381 cggtggctgt atttccagga gaaatagggg acgaggagac tggaatggag gatgagaccc
     4441 accgtgtcat cgatccattg gtggtcgctg aaggcgtggt ggccgcattc gtgggctatg
     4501 acccaaactc cggtgagcac acagccttgc gccacccagt acactggcca cgcgactgag
     4561 gagaggggtg gtgggagaag gtggaagtag gtggtagcga ggtggtagag tagaaaggcg
     4621 gctgtgaggt cgtatgcgac gtaggagaag gagcggagaa gtgagcggtt gaaacagtgg
     4681 ggagggattg cttttttaat gtcgccgacg gtgaaagggg gttttgtaga agggactctt
     4741 tcgagggggt tcctggtttt tctgacagct ggcatctgcc ctcctgctcc catattggat
     4801 ccggggtttt ttctccttga cgttaaagta tagaggtata ttaacaattt tttgttgata
     4861 cttttattac atttgaataa gaagtaatac aaaccgaaaa tgttgaaagt attagttaaa
     4921 gtggttatgc agtttttgca tttatatatc tgttaataga tcaaaaatca tcgcttcgct
     4981 gattaattac cccagaaata aggctaaaaa actaatcgca ttatcatcct atggttgtta
     5041 atttgattcg ttcatttgaa ggtttgtggg gccaggttac tgccaatttt tcctcttcat
     5101 aaccataaaa gctagtattg tagaatcttt attgttcgga gcagtgcggc gcgaggcaca
     5161 tctgcgtttc aggaacgcga ccggtgaaga cgaggacgca cggaggagag tcttccttcg
     5221 gagggctgtc acccgctcgg cggcttctaa tccgtacttc aatatagcaa tgagcagtta
     5281 agcgtattac tgaaagttcc aaagagaagg tttttttagg ctaagataat ggggctcttt
     5341 acatttccac aacatataag taagattaga tatggatatg tatatggata tgtatatggt
     5401 ggtaatgcca tgtaatatga ttattaaact tctttgcgtc catccaaaaa aaaagtaaga
     5461 atttttgaaa attcgaattc aaccctcact aaagggcggc cgcactagta tgggtggcgg
     5521 tggccgtggt cggacttcgc aaaaacccct catggaacgt gtctcagttg atccaccctt
     5581 caccgtgagt gatctcaagc aagcaatccc tccccattgc ttcaagcgat ctgtaatccg
     5641 ttcctcttac tacatagtcc acgatgctat tatcgcctac atcttctact tccttgccga
     5701 caaatacatt ccgattctcc ctgcccctct agcctacctc gcttggcccc tttactggtt
     5761 ctgtcaagct agcatcctca ccggcttatg ggtcatcggt cacgaatgcg gtcaccatgc
     5821 cttcagcgac taccagtggg ttgacgacac tgtgggcttc atcctccact cgtttctcat
     5881 gaccccgtat ttctcctgga aatacagcca ccggaaccac catgccaaca caaattcgct
     5941 tgacaacgat gaagtttaca tccccaaaag caaggccaaa gtcgcgcttt actataaagt
     6001 tctcaaccac ccacctggcc gactgttgat tatgttcatc accttcaccc taggcttccc
     6061 tctatacctc tttaccaata tttccggcaa gaagtatgaa aggtttgcca accatttcga
     6121 ccccatgagt ccgattttca aagagcgtga gcggtttcag gtcttgctat cggatcttgg
     6181 ccttcttgct gtgctttacg gagttaaact tgcggtagca gcgaaaggcg ccgcttgggt
     6241 gacgtgcatt tacggaattc cagttttagg cgtgtttatc tttttcgata tcatcaccta
     6301 cttgcaccac acccatctgt cgttgcctca ttatgattca tctgaatgga actggctcag
     6361 aggggctttg tcaacaatcg atagggactt tgggttcctg aatagtgtgc tccatgatgt
     6421 tacacacact cacgttatgc atcatctgtt ttcatacatt ccacactatc atgcgaagga
     6481 ggcaagggat gcaatcaaca cagtcttggg cgacttttat aagatcgata ggactccaat
     6541 tctgaaagca atgtggagag aggccaagga atgcatcttc atcgagcctg aaaaaggtag
     6601 ggagtccaag ggtgtatatt ggtacaataa attcgcacta gtatcgatgg attacaagga
     6661 tgacgacgat aagatctgag ctcttaatta acaattcttc gccagaggtt tggtcaagtc
     6721 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     6781 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     6841 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     6901 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     6961 ttctcaggta tagcatgagg tcgctccaat tcagctggcg taatagcgaa gaggcccgca
     7021 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     7081 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     7141 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7201 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7261 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7321 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7381 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7441 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     7501 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7561 tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     7621 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     7681 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     7741 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     7801 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     7861 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     7921 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     7981 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     8041 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     8101 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     8161 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     8221 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     8281 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     8341 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     8401 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     8461 tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     8521 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     8581 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaagaaacc gaaatcaaaa
     8641 aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct gtggtatggt gcactctcag
     8701 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     8761 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     8821 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
