LOCUS       sequence-129503-        6951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces Crepis alpina delta15 fatty acid desaturase (FAD3) in 
            yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6951)
  AUTHORS   Nam JW, Kappock TJ
  TITLE     Cloning and transcriptional analysis of Crepis alpina fatty acid 
            desaturases affecting the biosynthesis of crepenynic acid.
  JOURNAL   J Exp Bot. 2007;58(6):1421-32. Epub 2007 Feb 27.
  PUBMED    17329262
REFERENCE   2  (bases 1 to 6951)
  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: "J Exp 
            Bot."; date: "2007"; volume: "58(6)"; pages: "1421-32. Epub 2007 Feb
            27"
COMMENT     Sequence Label: pJK277
FEATURES             Location/Qualifiers
     source          1..6951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(1952..2755)
                     /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(2756..2976)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      3575..4455
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(4524..4979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4873..4892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4998..5440
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4998..5115
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5411..5434
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     5472..5491
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5472..5490
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5655..5666)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      6700..6947
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(6716..6734)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa gcccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggagcg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc attcagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttggcattgc tacaggcatc
     1321 gtggtgtcac tctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aatagtgtat cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     1921 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     1981 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     2041 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     2101 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     2161 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     2221 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     2281 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     2341 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     2401 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     2461 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     2521 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     2581 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     2641 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     2701 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     2761 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     2821 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     2881 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaagaaac cgaaatcaaa
     2941 aaaaagaata aaaaaaaaat gatgaattga attgaaaagc tagcttatcg atgataagct
     3001 gtcaaagatg agaattaatt ccacggacta tagactatac tagatactcc gtctactgta
     3061 cgatacactt ccgctcaggt ccttgtcctt taacgaggcc ttaccactct tttgttactc
     3121 tattgatcca gctcagcaaa ggcagtgtga tctaagattc tatcttcgcg atgtagtaaa
     3181 actagctaga ccgagaaaga gactagaaat gcaaaaggca cttctacaat ggctgccatc
     3241 attattatcc gatgtgacgc tgcagcttct caatgatatt cgaatacgct ttgaggagat
     3301 acagcctaat atccgacaaa ctgttttaca gatttacgat cgtacttgtt acccatcatt
     3361 gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat ttgaacctgt
     3421 ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg ttcctggaga
     3481 aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc attttctgcg
     3541 tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc ttcattttgt
     3601 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     3661 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     3721 tttgtaaaac aaaaatgcaa cgcgacgaga gcgctaattt ttcaaacaaa gaatctgagc
     3781 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     3841 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     3901 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     3961 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     4021 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     4081 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     4141 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     4201 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     4261 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     4321 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     4381 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     4441 agcggtattc gcaatgggaa gctccacccc ggttgataat cagaaaagcc ccaaaaacag
     4501 gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa aattcgcgtt
     4561 aaatttttgt taaatcagct cattttttaa cgaatagccc gaaatcggca aaatccctta
     4621 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttcca acaagagtcc
     4681 actattaaag aacgtggact ccaacgtcaa agggcgaaaa agggtctatc agggcgatgg
     4741 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcagt
     4801 aaatcggaag ggtaaacgga tgcccccatt tagagcttga cggggaaagc cggcgaacgt
     4861 ggcgagaaag gaagggaaga aagcgaaagg agcgggggct agggcggtgg gaagtgtagg
     4921 ggtcacgctg ggcgtaacca ccacacccgc cgcgcttaat ggggcgctac agggcgcgtg
     4981 gggatgatcc actagtacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag
     5041 actctcctcc gtgcgtcctc gtcctcaccg gtcgcgttcc tgaaacgcag atgtgcctcg
     5101 cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg
     5161 aaaaattggc agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca
     5221 taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg
     5281 atgatttttg atctattaac agatatataa atgcaaaaac tgcattaacc actttaacta
     5341 atactttcaa cattttcggt ttgtattact tcttattcaa atgtaataaa agtatcaaca
     5401 aaaaattgtt aatatacctc tatactttaa cgtcaaggag aaaaaacccc ggatcggact
     5461 actagcagct gtaatacgac tcactatagg gaatattaag cttggtaccg agctcggatc
     5521 cacaatggcg gtttcttctc cagatgccac agaaaagctt aatcaagctt cgcctctcgt
     5581 caatggcgtc caaaagaatc aagaagaatt cgacccaagt gcccctcctc cattcaaggt
     5641 ggctgacatc agagctgcaa ttcccagcca ttgctgggtc aagaatccat ggaggtctct
     5701 cagctacgtc ctcagggaca ttctggtaat ttcctcattg gtcgccatcg ctgttatctt
     5761 caaaaccagt tcatgggttt ggccgattta ctggattgca caaggcacaa tgttctgggc
     5821 aatctttgtt cttggtcatg attgtggcca tggaagcttt tcagacaacc ctaatctcaa
     5881 tagcgtcgtg ggccacattt tgcattcttc gattctcgta ccctaccatg ggtggagaat
     5941 cagccacagg acccatcatc aaaatcatgg tcatgtcgaa aatgatgaat catgggttcc
     6001 gttgaccgaa aagacttaca acacattgga tgcttccacc aaactactca gattcaaagt
     6061 cccttttccc ttatttgcct acccattgta cttgtggtat aggagcccag gcaaatcagg
     6121 gtcacatttc aacccgtata gtgatatgtt ctcccccaat gaaagaagtt atattgtgac
     6181 atctacttta tgttggtctc taatggtagc caatctggta tttttgtcaa ttgttattgg
     6241 cccaactcta ttgttcaacc tctatggcat tccttatttg atattcgtta tgtggttgga
     6301 ttttgtcact tatttgcatc accacggcca tgagcaaaaa cttccttggt atcgtggcaa
     6361 agaatggagc tacctaaggg gaggattaac aaccgttgat cgagattatg gaatattcaa
     6421 taacatccat catgacatag gcacccatgt catccatcat cttttcccac aaatcccaca
     6481 ttatcatctg attgaagcga caatggcggc aaagtctgtg cttgggaatt actaccgaga
     6541 gccaaaaaag tctggctgga taccggttca cttaatggat aacttggtga aaagcgtcaa
     6601 acaagatcat tatgttagtg atgcgggtga agtcgtttac tatcaaactg attatcgaat
     6661 gattgggaag aaaattgagt gattttctag agggccgcat catgtaatta gttatgtcac
     6721 gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga gttagacaac
     6781 ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga acgttattta
     6841 tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac attatactga
     6901 aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcggc c
//
