LOCUS Exported 5674 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Characterisation of TEF1 promoter using yEGFP as reporter gene and PGK1 terminator. ACCESSION . VERSION . KEYWORDS pILGFP1F5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5674) TITLE Vickers lab yeast integrative expression plasmids REFERENCE 2 (bases 1 to 5674) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5674 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 223..630 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 653..1363 /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" terminator 1373..1643 /gene="S. cerevisae PGK1" /label=PGK1 terminator /note="transcription terminator for 3-phosphoglycerate kinase gene" primer_bind complement(1908..1924) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1908..1924) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1921..1943) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1932..1948 /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(1956..1986) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2001..2022 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2139..2156) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2310..2898) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2390..2409) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3069..3929) /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 3692..3711 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3930..4034) /gene="bla" /label=AmpR promoter primer_bind 4102..4120 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4158..4180) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4280..4299 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4493..4515 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4507..4524 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4508..4524 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 5638..5671 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 aagggttgct cgagaaagag ctccagaaag cgaccaccca actttggctg ataatagcgt 61 ataaacaatg catactttgt acgttcaaaa tacaatgcag tagatatatt tatgcatatt 121 acatataata catatcacat aggaagcaac aggcgcgttg gacttttaat tttcgaggac 181 cgcgaatcct tacatcacac ccaatccccc acaagtgatc ccccacacac catagcttca 241 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac 301 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc 361 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct 421 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt 481 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat 541 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat 601 tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaaatgggat cctctaaagg 661 tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg 721 tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt 781 aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt 841 cggttatggt gttcaatgtt ttgcgagata cccagatcat atgaaacaac atgacttttt 901 caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg 961 taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga 1021 attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa 1081 ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa 1141 cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca 1201 aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact tatccactca 1261 atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt 1321 tactgctgct ggtattaccc atggtatgga tgaattgtac aaaagatctt aaattgaatt 1381 gaattgaaat cgatagatca atttttttct tttctctttc cccatccttt acgctaaaat 1441 aatagtttat tttatttttt gaatattttt tatttatata cgtatatata gactattatt 1501 tatcttttaa tgattattaa gatttttatt aaaaaaaaat tcgctcctct tttaatgcct 1561 ttatgcagtt tttttttccc attcgatatt tctatgttcg ggttcagcgt attttaagtt 1621 taataactcg aaaattctgc gttcgttaaa gctttcgaga aggatattat ttactagtag 1681 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct 1741 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg 1801 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccggga 1861 atctcggtcg taatgattta aatgcatgca agcttggcgt aatcatggtc atagctgttt 1921 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 1981 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 2041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 2101 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 2161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 2221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 2281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 2401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 2461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 2521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 2581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 2641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 2701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 2761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 2821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 2881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 2941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 3121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 3181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 3241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 3301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 3361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 3421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 3481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 3541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 3601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 3661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 3721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 3781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 3841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 3901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 3961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 4021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 4081 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 4141 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 4201 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 4261 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 4321 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 4381 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 4441 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca 4501 cgacgttgta aaacgacggc cagtgaattc atttaaattt ttttgattcg gtaatctccg 4561 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta 4621 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct 4681 gcaggaaacg aagataaaac atgtccacaa aatcatatac cagtagagct gagactcatg 4741 caagtccggt tgcatcgaaa cttttacgtt taatggatga aaagaagacc aatttgtgtg 4801 cttctcttga cgttcgttcg actgatgagc tattgaaact tgttgaaacg ttgggtccat 4861 acatttgcct tttgaaaaca cacgttgata tcttggatga tttcagttat gagggtactg 4921 tcgttccatt gaaagcattg gcagagaaat acaagttctt gatatttgag gacagaaaat 4981 tcgccgatat cggtaacaca gtcaaattac aatatacatc gggcgtttac cgtatcgcag 5041 aatggtctga tatcaccaac gcccacgggg ttactggtgc tggtattgtt gctggcttga 5101 aacaaggtgc gcaagaggtc accaaagaac caaggggatt attgatgctt gctgaattgt 5161 cttccaaggg ttctctagca cacggtgaat atactaaggg taccgttgat attgcaaaga 5221 gtgataaaga tttcgttatt gggttcattg ctcagaacga tatgggagga agagaagaag 5281 ggtttgattg gctaatcatg accccaggtg taggtttaga cgacaaaggc gatgcattgg 5341 gtcagcagta cagaaccgtc gacgaagttg taagtggtgg atcagatatc atcattgttg 5401 gcagaggact tttcgccaag ggtagagatc ctaaggttga aggtgaaaga tacagaaatg 5461 ctggatggga agcgtaccaa aagagaatca gcgctcccca ttaattatac aggaaactta 5521 atagaacaaa tcacatattt aatctaatag ccacctgcat tggcacggtg caacactact 5581 tcaacttcat cttacaaaaa gatcacgtga tctgttgtat tgggatctct agacctaata 5641 acttcgtata gcatacatta tacgaagtta tatt //