LOCUS       sequence-136122-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Crepis alpina delta-12 fatty acid acetylenase (vFAD2) with 
            C-terminal FLAG epitope.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Kappock
  TITLE     Additional fatty acid desaturase and acetylenase constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7764)
  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            "Unpublished"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: pJK369
FEATURES             Location/Qualifiers
     source          1..7764
                     /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"
     primer_bind     complement(3670..3689)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3671..3689)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3700..4364
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(3713..3736)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4031..4148
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             5544..5567
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      5715..5880
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(5770..5794)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      5959..6414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6046..6065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6256..6277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6545..7348)
                     /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(7349..7569)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7595..7614)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7714..7736
                     /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 acgcccgggc cctatagtga gtcgtattac ggatccgggg ttttttctcc ttgacgttaa
     3721 agtatagagg tatattaaca attttttgtt gatactttta ttacatttga ataagaagta
     3781 atacaaaccg aaaatgttga aagtattagt taaagtggtt atgcagtttt tgcatttata
     3841 tatctgttaa tagatcaaaa atcatcgctt cgctgattaa ttaccccaga aataaggcta
     3901 aaaaactaat cgcattatca tcctatggtt gttaatttga ttcgttcatt tgaaggtttg
     3961 tggggccagg ttactgccaa tttttcctct tcataaccat aaaagctagt attgtagaat
     4021 ctttattgtt cggagcagtg cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg
     4081 aagacgagga cgcacggagg agagtcttcc ttcggagggc tgtcacccgc tcggcggctt
     4141 ctaatccgta cttcaatata gcaatgagca gttaagcgta ttactgaaag ttccaaagag
     4201 aaggtttttt taggctaaga taatggggct ctttacattt ccacaacata taagtaagat
     4261 tagatatgga tatgtatatg gatatgtata tggtggtaat gccatgtaat atgattatta
     4321 aacttctttg cgtccatcca aaaaaaaagt aagaattttt gaaaattcga attcaaccct
     4381 cactaaaggg cggccgcact agtatgggtg gcggtggccg tggtcggact tcgcaaaaac
     4441 ccctcatgga acgtgtctca gttgatccac ccttcaccgt gagtgatctc aagcaagcaa
     4501 tccctcccca ttgcttcaag cgatctgtaa tccgttcctc ttactacata gtccacgatg
     4561 ctattatcgc ctacatcttc tacttccttg ccgacaaata cattccgatt ctccctgccc
     4621 ctctagccta cctcgcttgg cccctttact ggttctgtca agctagcatc ctcaccggct
     4681 tatgggtcat cggtcacgaa tgcggtcacc atgccttcag cgactaccag tgggttgacg
     4741 acactgtggg cttcatcctc cactcgtttc tcatgacccc gtatttctcc tggaaataca
     4801 gccaccggaa ccaccatgcc aacacaaatt cgcttgacaa cgatgaagtt tacatcccca
     4861 aaagcaaggc caaagtcgcg ctttactata aagttctcaa ccacccacct ggccgactgt
     4921 tgattatgtt catcaccttc accctaggct tccctctata cctctttacc aatatttccg
     4981 gcaagaagta tgaaaggttt gccaaccatt tcgaccccat gagtccgatt ttcaaagagc
     5041 gtgagcggtt tcaggtcttg ctatcggatc ttggccttct tgctgtgctt tacggagtta
     5101 aacttgcggt agcagcgaaa ggcgccgctt gggtgacgtg catttacgga attccagttt
     5161 taggcgtgtt tatctttttc gatatcatca cctacttgca ccacacccat ctgtcgttgc
     5221 ctcattatga ttcatctgaa tggaactggc tcagaggggc tttgtcaaca atcgataggg
     5281 actttgggtt cctgaatagt gtgctccatg atgttacaca cactcacgtt atgcatcatc
     5341 tgttttcata cattccacac tatcatgcga aggaggcaag ggatgcaatc aacacagtct
     5401 tgggcgactt ttataagatc gataggactc caattctgaa agcaatgtgg agagaggcca
     5461 aggaatgcat cttcatcgag cctgaaaaag gtagggagtc caagggtgta tattggtaca
     5521 ataaattcgc actagtatcg atggattaca aggatgacga cgataagatc tgagctctta
     5581 attaacaatt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     5641 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     5701 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     5761 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttat
     5821 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     5881 caattcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5941 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6001 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6061 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6121 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6181 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6241 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6301 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6361 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat
     6421 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     6481 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     6541 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     6601 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     6661 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     6721 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     6781 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     6841 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     6901 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     6961 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     7021 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     7081 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     7141 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     7201 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     7261 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     7321 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     7381 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     7441 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     7501 aatcaaaaaa atttcaaaga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     7561 tgaattgaaa agctgtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7621 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7681 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7741 caccgtcatc accgaaacgc gcga
//
