LOCUS       sequence-96592-e        6435 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Contains a promoter comprised of derivatives from native promoters 
            galactose, cyc and leucine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6435)
  AUTHORS   Blazeck J, Garg R, Reed B, Alper HS
  TITLE     Controlling promoter strength and regulation in Saccharomyces 
            cerevisiae using synthetic hybrid promoters.
  JOURNAL   Biotechnol Bioeng. 2012 Nov;109(11):2884-95. doi: 10.1002/bit.24552.
            Epub 2012 May 17.
  PUBMED    22565375
REFERENCE   2  (bases 1 to 6435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1002/bit.24552";
            journalName: "Biotechnol Bioeng"; date: "2012-11"; volume: "109"; 
            issue: "11"; pages: "2884-95"
COMMENT     Sequence Label: UASGAL-CU2-Pleum
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..153
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(306..311,312..329,330..336)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    344..360
                     /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)."
     primer_bind     349..371
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     368..384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     368..384
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     403..423
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        405..423
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    475..592
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             1101..1817
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1820..1836)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1821..1837)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1855..1873)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2087..2106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2088..2106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2116..2133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2116..2132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2125..2147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2277..2732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2364..2383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2574..2595
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2863..3666)
                     /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(3667..3887)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3913..3932)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4032..4054
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4093..4111)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4153..4656
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4693..4797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4798..4866,4867..5658)
                     /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(5016..5035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5829..6417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6318..6337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
       61 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      121 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
      301 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
      361 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
      421 agggaacaaa agctggagct ccgggatcct tggcgcgccc cttaattaat agtacggatt
      481 agaagccgcc gagcgggcga cagccctccg acggaagact ctcctccgtg cgtcctcgtc
      541 ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa
      601 agattctaca atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc
      661 cacaaacctt caaattaacg aatcaaatta acaaccatag gatgataatg cgattagttt
      721 tttagcctta tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga
      781 agcttatttg gcgagcgttg gttggtggat caagcccacg cgtaggcaat cctcgagcag
      841 atccgccagg cgtgtatata tagcgtggat ggccaggcaa ctttagtgct gacacataca
      901 ggcatatata tatgtgtgcg acgacacatg atcatatggc atgcatgtgc tctgtatggt
      961 ttaaaccaat attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt
     1021 actttctaac ttttcttacc ttttacattt cagcaatata tatatatata tttcaaggat
     1081 ataccattct atctagaaaa atgtctaaag gtgaagaatt attcactggt gttgtcccaa
     1141 ttttggttga attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg
     1201 aaggtgatgc tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc
     1261 cagttccatg gccaacctta gtcactactt taggttatgg tttgatgtgt tttgctagat
     1321 acccagatca tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc
     1381 aagaaagaac tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt
     1441 ttgaaggtga taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg
     1501 gtaacatttt aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcatgg
     1561 ctgacaaaca aaagaatggt atcaaagtta acttcaaaat tagacacaac attgaagatg
     1621 gttctgttca attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct
     1681 tgttaccaga caaccattac ttatcctatc aatctgcctt atccaaagat ccaaacgaaa
     1741 agagagacca catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg
     1801 atgaattgta caaataaatc gataccgtcg acctcgagtc atgtaattag ttatgtcacg
     1861 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     1921 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     1981 atatttcaaa tttttctttt ttttctgtac aaacgcgtgt acgcatgtaa cattatactg
     2041 aaaaccttgc ttgagaaggt tttgggacgc tcgggtaccc aattcgccct atagtgagtc
     2101 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc
     2281 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2341 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2401 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2461 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2521 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2581 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2641 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2701 gaattttaac aaaatattaa cgtttacaat ttcctgatgc ggtattttct ccttacgcat
     2761 ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg aagctctaat
     2821 ttgtgagttt agtatacatg catttactta taatacagtt ttttagtttt gctggccgca
     2881 tcttctcaaa tatgcttccc agcctgcttt tctgtaacgt tcaccctcta ccttagcatc
     2941 ccttcccttt gcaaatagtc ctcttccaac aataataatg tcagatcctg tagagaccac
     3001 atcatccacg gttctatact gttgacccaa tgcgtctccc ttgtcatcta aacccacacc
     3061 gggtgtcata atcaaccaat cgtaaccttc atctcttcca cccatgtctc tttgagcaat
     3121 aaagccgata acaaaatctt tgtcgctctt cgcaatgtca acagtaccct tagtatattc
     3181 tccagtagat agggagccct tgcatgacaa ttctgctaac atcaaaaggc ctctaggttc
     3241 ctttgttact tcttctgccg cctgcttcaa accgctaaca atacctgggc ccaccacacc
     3301 gtgtgcattc gtaatgtctg cccattctgc tattctgtat acacccgcag agtactgcaa
     3361 tttgactgta ttaccaatgt cagcaaattt tctgtcttcg aagagtaaaa aattgtactt
     3421 ggcggataat gcctttagcg gcttaactgt gccctccatg gaaaaatcag tcaagatatc
     3481 cacatgtgtt tttagtaaac aaattttggg acctaatgct tcaactaact ccagtaattc
     3541 cttggtggta cgaacatcca atgaagcaca caagtttgtt tgcttttcgt gcatgatatt
     3601 aaatagcttg gcagcaacag gactaggatg agtagcagca cgttccttat atgtagcttt
     3661 cgacatgatt tatcttcgtt tcctgcaggt ttttgttctg tgcagttggg ttaagaatac
     3721 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
     3781 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaagaaa
     3841 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aattgaaaag ctgtggtatg
     3901 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3961 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4021 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4081 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4141 tttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga
     4201 ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa
     4261 agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa
     4321 aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa
     4381 atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg
     4441 tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag
     4501 ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa
     4561 actatttttt ctttaatttc tttttttact ttctattttt aatttatata tttatattaa
     4621 aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg tggcactttt
     4681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5041 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5341 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5941 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6421 gccagcaacg cggcc
//
