LOCUS       sequence-138462-        8886 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 (FAD2-2) 
            with C-terminal c-myc epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8886)
  AUTHORS   Kappock
  TITLE     Additional fatty acid desaturase and acetylenase constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8886)
  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            "Unpublished"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: pJK370
FEATURES             Location/Qualifiers
     source          1..8886
                     /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        4822..5486
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(4835..4858)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5153..5270
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             6666..6689
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6837..7002
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(6892..6916)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      7081..7536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7168..7187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7378..7399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7667..8470)
                     /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(8471..8691)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(8717..8736)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8836..8858
                     /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 agtatcttat tcttgtacca aaacacaccg cccttttcct caccatccgg ctccacaaac
     3721 aaacactctt tggcctccct ccacatggca acataccacg gcgtctcatc caagtgataa
     3781 tactccccaa gcaccggcct caccgccttc gtcgcctcca tcgcgttata atgaggcatc
     3841 gttgaaaaca aatggtgcgc cacatgcgtg tctgtaatat tatggaaaac cttattcaaa
     3901 acgccgtagt ccctatccac cgtcgccagc gcccctctca gccaatccca ctccgacccg
     3961 tcgtaatgag gcaacgacgg gtgtgtatgt tgcaggtaag taatcaaaac cagaaatccg
     4021 ttcacaatca gtaacggaac gccgtaaacg caaatcaccc aactcacccc tttcagaacg
     4081 gcgaggcggt acaggaagta agacatggtg gcgattccga cgtcggagag ccagatctgg
     4141 aggcgttcac ggtcgttgta gatagggctg ttggggacgt agtgggaggc gaagcggtcg
     4201 tacggtctgc cggagacgtt gaacgctagg tataacggcc agccgagagt gagagtcgcg
     4261 agcatgctta ttatccggcc gggtgggttg tttaggtatt ttgagtacca agtgatttta
     4321 gatctgggtt tcgggacgaa cacttcgtcg cggtcgagtg agccggtgtt ggagtggtgg
     4381 cggcggtggc tgtatttcca tgagaagtag gggacgagga gggcggagtg gaggaggaat
     4441 ccgacggtgt cgtcgacgaa ttggtagtcg ctgaatgcgt ggtggccgca ttcgtgggcg
     4501 atgacccaga cgccggtgag gacgcagcct tgggtgatcc agtaagggag ccaggcgagg
     4561 gaggagagag gcggagggag gtggtggaag taggcggtgg cgatgtggta gaggaggaag
     4621 ctgatgatga gatcggaaac gacgtaggag aaggaacggg tgagggagcg gttgaaacag
     4681 tggggaggga tgactttttt gaggtcgctg attgtgaatg gaggtttttc agaagggact
     4741 cgtttcagag ggttgttttt ttgttcggat tttgtatcgg aaacagacat acatcctcct
     4801 gcccccattt ttggatccgg ggttttttct ccttgacgtt aaagtataga ggtatattaa
     4861 caattttttg ttgatacttt tattacattt gaataagaag taatacaaac cgaaaatgtt
     4921 gaaagtatta gttaaagtgg ttatgcagtt tttgcattta tatatctgtt aatagatcaa
     4981 aaatcatcgc ttcgctgatt aattacccca gaaataaggc taaaaaacta atcgcattat
     5041 catcctatgg ttgttaattt gattcgttca tttgaaggtt tgtggggcca ggttactgcc
     5101 aatttttcct cttcataacc ataaaagcta gtattgtaga atctttattg ttcggagcag
     5161 tgcggcgcga ggcacatctg cgtttcagga acgcgaccgg tgaagacgag gacgcacgga
     5221 ggagagtctt ccttcggagg gctgtcaccc gctcggcggc ttctaatccg tacttcaata
     5281 tagcaatgag cagttaagcg tattactgaa agttccaaag agaaggtttt tttaggctaa
     5341 gataatgggg ctctttacat ttccacaaca tataagtaag attagatatg gatatgtata
     5401 tggatatgta tatggtggta atgccatgta atatgattat taaacttctt tgcgtccatc
     5461 caaaaaaaaa gtaagaattt ttgaaaattc gaattcaacc ctcactaaag ggcggccgca
     5521 ctagtatggg tggcggtggc cgtggtcgga cttcgcaaaa acccctcatg gaacgtgtct
     5581 cagttgatcc acccttcacc gtgagtgatc tcaagcaagc aatccctccc cattgcttca
     5641 agcgatctgt aatccgttcc tcttactaca tagtccacga tgctattatc gcctacatct
     5701 tctacttcct tgccgacaaa tacattccga ttctccctgc ccctctagcc tacctcgctt
     5761 ggccccttta ctggttctgt caagctagca tcctcaccgg cttatgggtc atcggtcacg
     5821 aatgcggtca ccatgccttc agcgactacc agtgggttga cgacactgtg ggcttcatcc
     5881 tccactcgtt tctcatgacc ccgtatttct cctggaaata cagccaccgg aaccaccatg
     5941 ccaacacaaa ttcgcttgac aacgatgaag tttacatccc caaaagcaag gccaaagtcg
     6001 cgctttacta taaagttctc aaccacccac ctggccgact gttgattatg ttcatcacct
     6061 tcaccctagg cttccctcta tacctcttta ccaatatttc cggcaagaag tatgaaaggt
     6121 ttgccaacca tttcgacccc atgagtccga ttttcaaaga gcgtgagcgg tttcaggtct
     6181 tgctatcgga tcttggcctt cttgctgtgc tttacggagt taaacttgcg gtagcagcga
     6241 aaggcgccgc ttgggtgacg tgcatttacg gaattccagt tttaggcgtg tttatctttt
     6301 tcgatatcat cacctacttg caccacaccc atctgtcgtt gcctcattat gattcatctg
     6361 aatggaactg gctcagaggg gctttgtcaa caatcgatag ggactttggg ttcctgaata
     6421 gtgtgctcca tgatgttaca cacactcacg ttatgcatca tctgttttca tacattccac
     6481 actatcatgc gaaggaggca agggatgcaa tcaacacagt cttgggcgac ttttataaga
     6541 tcgataggac tccaattctg aaagcaatgt ggagagaggc caaggaatgc atcttcatcg
     6601 agcctgaaaa aggtagggag tccaagggtg tatattggta caataaattc gcactagtat
     6661 cgatggatta caaggatgac gacgataaga tctgagctct taattaacaa ttcttcgcca
     6721 gaggtttggt caagtctcca atcaaggttg tcggcttgtc taccttgcca gaaatttacg
     6781 aaaagatgga aaagggtcaa atcgttggta gatacgttgt tgacacttct aaataagcga
     6841 atttcttatg atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca
     6901 aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc
     6961 tgtaggtcag gttgctttct caggtatagc atgaggtcgc tccaattcag ctggcgtaat
     7021 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     7081 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     7141 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     7201 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     7261 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     7321 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     7381 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     7441 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     7501 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     7561 gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc
     7621 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     7681 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     7741 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     7801 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     7861 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     7921 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     7981 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     8041 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     8101 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     8161 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     8221 acttggcgga taatgccttt agcggcttaa ctgtgccctc catggaaaaa tcagtcaaga
     8281 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     8341 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     8401 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     8461 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     8521 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     8581 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaaa
     8641 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaattga aaagctgtgg
     8701 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     8761 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     8821 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     8881 gcgcga
//
