LOCUS       sequence-168933-        5476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express yEGFP-ERG9 ER signal peptide under the control of TEF1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  AUTHORS   Peng B, Plan MR, Chrysanthopoulos P, Hodson MP, Nielsen LK, Vickers 
            CE
  TITLE     A squalene synthase protein degradation method for improved 
            sesquiterpene production in Saccharomyces cerevisiae.
  JOURNAL   Metab Eng. 2017 Jan;39:209-219. doi: 10.1016/j.ymben.2016.12.003. 
            Epub 2016 Dec 8.
  PUBMED    27939849
REFERENCE   2  (bases 1 to 5476)
  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: "Metab 
            Eng."; date: "2017-01"; volume: "39:209-219. doi"; pages: " 
            10.1016/j.ymben.2016.12.003. Epub 2016 Dec 8"
COMMENT     Sequence Label: pILGFPJ1
FEATURES             Location/Qualifiers
     source          1..5476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..716
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1063..1079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1063..1079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1076..1098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1087..1103
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1111..1117,1118..1135,1136..1141))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1156..1177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1294..1311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1465..2053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1545..1564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2224..3015,3016..3084))
                     /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     2847..2866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3085..3189)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3257..3275
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3313..3335)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3435..3454
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3648..3670
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3662..3679
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3663..3679
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4793..4826)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5037..5450
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 gatcctctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg
       61 gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg
      121 gtaaattgac cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct
      181 tagtcactac tttcggttat ggtgttcaat gttttgcgag atacccagat catatgaaac
      241 aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt
      301 tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag
      361 ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca
      421 aattggaata caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg
      481 gtatcaaagt taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg
      541 accattatca acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt
      601 acttatccac tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct
      661 tgttagaatt tgttactgct gctggtatta cccatggtat ggatgaattg tacaaaagat
      721 ccgttccaac ccaacaagaa gaagagtaca agttcaatat ggttttatct atcatcttgt
      781 ccgttcttct tgggttttat tatatataca ctttacacag agcgtgaact agtaggacta
      841 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
      901 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
      961 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgggaatctc
     1021 ggtcgtaatg atttaaatgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     1081 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1141 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1201 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     1261 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1321 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1381 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1441 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1501 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1561 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     1621 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     1681 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     1741 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     1801 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     1861 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     1921 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     1981 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2041 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2101 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2161 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2221 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2281 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2341 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     2401 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2461 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2521 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     2581 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     2641 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2701 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     2761 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2821 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2881 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2941 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3001 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3061 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3121 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3181 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3241 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     3301 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     3361 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     3421 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     3481 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     3541 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     3601 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     3661 ttgtaaaacg acggccagtg aattcattta aatttttttg attcggtaat ctccgagcag
     3721 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtggtgtt
     3781 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
     3841 aaacgaagat aaaacatgtc cacaaaatca tataccagta gagctgagac tcatgcaagt
     3901 ccggttgcat cgaaactttt acgtttaatg gatgaaaaga agaccaattt gtgtgcttct
     3961 cttgacgttc gttcgactga tgagctattg aaacttgttg aaacgttggg tccatacatt
     4021 tgccttttga aaacacacgt tgatatcttg gatgatttca gttatgaggg tactgtcgtt
     4081 ccattgaaag cattggcaga gaaatacaag ttcttgatat ttgaggacag aaaattcgcc
     4141 gatatcggta acacagtcaa attacaatat acatcgggcg tttaccgtat cgcagaatgg
     4201 tctgatatca ccaacgccca cggggttact ggtgctggta ttgttgctgg cttgaaacaa
     4261 ggtgcgcaag aggtcaccaa agaaccaagg ggattattga tgcttgctga attgtcttcc
     4321 aagggttctc tagcacacgg tgaatatact aagggtaccg ttgatattgc aaagagtgat
     4381 aaagatttcg ttattgggtt cattgctcag aacgatatgg gaggaagaga agaagggttt
     4441 gattggctaa tcatgacccc aggtgtaggt ttagacgaca aaggcgatgc attgggtcag
     4501 cagtacagaa ccgtcgacga agttgtaagt ggtggatcag atatcatcat tgttggcaga
     4561 ggacttttcg ccaagggtag agatcctaag gttgaaggtg aaagatacag aaatgctgga
     4621 tgggaagcgt accaaaagag aatcagcgct ccccattaat tatacaggaa acttaataga
     4681 acaaatcaca tatttaatct aatagccacc tgcattggca cggtgcaaca ctacttcaac
     4741 ttcatcttac aaaaagatca cgtgatctgt tgtattggga tctctagacc taataacttc
     4801 gtatagcata cattatacga agttatatta agggttgctc gaccagaaag cgaccaccca
     4861 actttggctg ataatagcgt ataaacaatg catactttgt acgttcaaaa tacaatgcag
     4921 tagatatatt tatgcatatt acatataata catatcacat aggaagcaac aggcgcgttg
     4981 gacttttaat tttcgaggac cgcgaatcct tacatcacac ccaatccccc acaagtgatc
     5041 ccccacacac catagcttca aaatgtttct actccttttt tactcttcca gattttctcg
     5101 gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct
     5161 ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag
     5221 accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt
     5281 ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa
     5341 gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac
     5401 tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ttttaattac
     5461 aaactcgaga aaatgg
//
