LOCUS       sequence-433677-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduction of a LowTempGAL Turbo module (HD-GAL3C) to accelerate 
            induction of GAL promoter induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7639)
  AUTHORS   Lu Z, Shen Q, Bandari NC, Evans S, McDonnell L, Liu L, Jin W, 
            Luna-Flores CH, Collier T, Talbo G, McCubbin T, Esquirol L, Myers C,
            Trau M, Dumsday G, Speight R, Howard CB, Vickers CE, Peng B
  TITLE     LowTempGAL: a highly responsive low temperature-inducible GAL system
            in Saccharomyces cerevisiae.
  JOURNAL   Nucleic Acids Res. 2024 May 29:gkae460. doi: 10.1093/nar/gkae460.
  PUBMED    38808673
REFERENCE   2  (bases 1 to 7639)
  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: "Nucleic 
            Acids Res. 2024 May 29:gkae460. doi: 10.1093/nar/gkae460."
COMMENT     Sequence Label: pIRHDGAL3C
FEATURES             Location/Qualifiers
     source          1..7639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(225..242)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(225..241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(234..256)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(450..469)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     569..591
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(629..647)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        715..819
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(820..888,889..1680)
                     /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(1038..1057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1851..2439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2340..2359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2593..2610
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2727..2748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2763..2768,2769..2786,2787..2793)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2801..2817
                     /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     2806..2828
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2825..2841
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2825..2841
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3218..3561
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      3954..4151
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             5016..5243
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             complement(5821..5838)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 cctatggatt ggtttgaaga ttttggtgag tagagcacgc agcacgctgt atttacgtat
       61 ttaattttat atatttgtgc atacactact agggaagact tgaaaaaaac ctaggaaatg
      121 aaaaaacgac acaggaagtc ccgtatttac tattttttcc ttccttttga tggggcaggg
      181 cggaaataga ggataggata agcctactgc ttagctgttt aaacactggc cgtcgtttta
      241 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
      301 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
      361 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      421 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      481 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
      541 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
      601 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      661 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      721 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      781 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      841 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      901 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      961 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1021 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1081 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1141 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1201 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1261 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1321 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1381 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1441 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1501 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1561 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1621 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1681 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1741 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1981 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2461 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2521 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2581 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2641 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2701 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2761 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2821 cacacaggaa acagctatga ccatgattac gccagtttaa actgtgggag attagagggc
     2881 aaagtgtaca aagataccac tttgtatacg acgctgtctc catgcgacat gtgtacaggt
     2941 gccatcatca tgtatggtat tccacgctgt gttgtcggtg agaacgttaa tttcaaaagt
     3001 aagggcgatt aattaaggat ccgctgaagc ttcgtacgct gcaggtcgac aacccttaat
     3061 gttcttcatt ctacgcacgg tccttcaatt cgcttggcgg agatctatga caagagggtc
     3121 gaactcgcct aagtcgtaat tgagtcagtt acgctaggga taacagggta atatagctgt
     3181 ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc agaataccct
     3241 ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc cgtacattta
     3301 gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc gcacggcgcg
     3361 aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc ccctcacaga
     3421 cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     3481 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     3541 cacatccgaa cataaacaac catgggtatg accgaccaag cgacgcccaa cctgccatca
     3601 cgagatttcg atcccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3661 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3721 gggctcgatc ccctcgcgag ttggttcagc tgctgcctga ggctggacga cctcgcggag
     3781 ttctaccggc agtgcaaatc cgtcggcatc caggaaacca gcagcggcta tccgcgcatc
     3841 catgcccccg aactgcagga gtggggaggc acgatggccg ctttggtcga cccggacggg
     3901 acgctcctgc gcctgataca gaacgaattg cttgcaggca tctcatgatc agtactgaca
     3961 ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg
     4021 ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc
     4081 cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg
     4141 tcgctatact gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa acgagctctc
     4201 gagaaccctt aaagttacgc tagggataac agggtaatat aggttcttca ttctacgcac
     4261 ggtccttcaa ttcgcttggc ggtctagaat gacaagaggg tcgaactcgc ctaagtcgta
     4321 attgagtcca gatccaccgc ggaagtgtct cataaacttt actcaagagt taagtactga
     4381 aaaattcgac ttttatgata gttcaagtgt cgacgctgcg ggtatagaaa gggttcttta
     4441 ctctatagta cctcctcgct cagcatctgc ttcttcccaa agatgaacgc ggcgttatgt
     4501 cactaacgac gtgcaccaac ttgcggaaag tggaatcccg ttccaaaact ggcatccact
     4561 aattgataca tctacacacc gcacgccttt tttctgaagc ccactttcgt ggactttgcc
     4621 atatgcaaaa ttcatgaagt gtgataccaa gtcagcatac acctcactag ggtagtttct
     4681 ttggttgtat tgatcatttg gttcatcgtg gttcattaat tttttttctc cattgctttc
     4741 tggctttgat cttactatca tttggatttt tgtcgaaggt tgtagaattg tatgtgacaa
     4801 gtggcaccaa gcatatataa aaaaaaaaag cattatcttc ctaccagagt tgattgttaa
     4861 aaacgtattt atagcaaacg caattgtaat taattcttat tttgtatctt ttcttccctt
     4921 gtctcaatct tttattttta ttttattttt cttttcttag tttctttcat aacaccaagc
     4981 aactaatact ataacataca ataataggta ccaaaatgca gattttcgtc aagactttga
     5041 ccggtaaaac cataacattg gaagttgaat cttccgatac catcgacaac gttaagtcga
     5101 aaattcaaga caaggaaggt atccctccag atcaacaaag attgatcttt gccggtaagc
     5161 agctagaaga cggtagaacg ctgtctgatt acaacattca gaaggagtcc accttacatc
     5221 ttgtgctaag gctaagaggt ggtagacatg gttctggtat catggttaga ccattgaact
     5281 gtatcgttgc tgtttctcaa aacatgggta ttggaaagaa tggtgacttg ccttggccac
     5341 cattgagaaa tgagtttaag tacttccaga gaatgaccac cacttcttca gttgaaggta
     5401 aacaaaactt ggtcatcatg ggtagaaaga cctggttttc tattccagaa aagaacaggt
     5461 tgttgaagga cagaatcaac atcgttttgt ccagggaatt gaaagaacca ccaagaggtg
     5521 ctcatttctt ggcaaaatct ttggatgatg ccttgagatt gatcgaacaa ccagaattgg
     5581 cttccaaggt tgatatggtt tggattgtcg gtggttcatc tgtttatcaa gaagccatga
     5641 atcaaccagg tcatttgagg ttgttcgtta ccagaatcat gcaagagttt gagtctgata
     5701 ctttcttccc tgaaattgac ttgggcaagt ataagttgtt gccagaatat ccaggtgtct
     5761 tgtctgaagt tcaagaagaa aagggtatca agtacaagtt cgaagtctac gaaaagaagg
     5821 atggtggtgg tggtggtggt aatacaaacg ttccaatatt cagttctccg gtcagagatt
     5881 taccaaggtc tttcgaacaa aaacatttag cggttgtaga tgcatttttc caaacatacc
     5941 atgtcaaacc tgattttatc gctaggtctc ctggcagagt aaatctgatt ggtgagcata
     6001 tagattattg cgatttttca gttttgccat tagccattga tgtggatatg ctttgcgcag
     6061 ttaaaatttt agacgaaaaa aatccatcca ttaccttaac aaatgcggac cctaaatttg
     6121 ctcagcgaaa gtttgatctg cctttagatg gttcctacat ggccatagat ccgtctgtgt
     6181 cggaatggtc gaattacttt aaatgcggac tacatgtggc acattcatac ttgaaaaaaa
     6241 ttgctccgga aagatttaat aatacaccct tagtaggtgc gcagatcttt tgccagagcg
     6301 atattcctac tggtggtgga ctctcatctg catttacttg cgcggcagca ctagccacaa
     6361 ttagagccaa tatgggaaaa aattttgata tttccaaaaa agacttgacc cgcatcacag
     6421 cggttgctga gcactatgtt ggagtcaata atggtggtat ggatcaagca acgtctgttt
     6481 atggggaaga agatcatgct ctatacgtag agtttaggcc aaaactaaag gccacacctt
     6541 tcaagtttcc tcaattgaaa aatcatgaaa tcagtttcgt catcgccaat actcttgtaa
     6601 agtctaataa attcgaaact gctcctacaa attacaattt aagagtaata gaggtaacag
     6661 ttgctgccaa cgccttggcg accagataca gcgtggcctt accatctcac aaggacaatt
     6721 ctaactcaga aagagggaat ctaagagatt ttatggatgc ttactacgcc agatacgaaa
     6781 accaagccca accatggaat ggagatatcg gaactggtat tgaacgttta ctcaagatgc
     6841 tacaattggt agaagaatct ttctcgagga aaaagagcgg attcactgta catgaagcct
     6901 ctacggcgct aaactgttca cgtgaggagt tcactagaga ttacctgaca acttttcccg
     6961 tccgcttcca agtcttgaaa ctatatcaaa gagctaaaca cgtttactcc gaatccttaa
     7021 gggtgcttaa ggctttaaaa atgatgacaa gtgccacttt tcacacggac gaagatttct
     7081 ttacagattt tggccgacta atgaatgagt cccaggcctc ttgtgataaa ctttatgaat
     7141 gttcgtgcat agaaaccaat caaatatgct cgattgccct agcaaatggt tctttcggct
     7201 cccgtctcac tggcgctggt tggggcggtt gcactatcca ccttgttcct agtggcgcta
     7261 atgggaacgt ggaacaggta cgaaaagcac taatcgaaaa attctacaat gtaagatatc
     7321 cggatctcac agatgaagaa ctaaaagacg caattatagt tccaaagcct gccttgggta
     7381 cttgtttgta cgaacaataa agtgctttta actaagaatt attagtcttt tctgcttatt
     7441 ttttcatcat agtttagaac actttatatt aacgaatagt ttatgaatct atttaggttt
     7501 aaaaattgat acagttttat aagttacttt ttcaaagact cgtgctgtct attgcataat
     7561 gcactggaag gggaaaaaaa aggtgcacac gcgtggcttt ttcttgaatt tgcagtttga
     7621 aaaataacta catgtaagg
//
