LOCUS       sequence-433678-        8135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduction of a LowTempGAL Turbo module (GAL4) to accelerate 
            induction of GAL promoter induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8135)
  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 8135)
  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: pIRGAL4A
FEATURES             Location/Qualifiers
     source          1..8135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             798..3443
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4"
                     /product="GAL4 transcriptional activator"
                     /label=GAL4
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     complement(4049..4066)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4049..4065)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4058..4080)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4274..4293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4393..4415
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4453..4471)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4539..4643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4644..4712,4713..5504)
                     /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(4862..4881)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5675..6263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6164..6183
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6417..6434
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6551..6572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6587..6592,6593..6610,6611..6617)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6625..6641
                     /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     6630..6652
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6649..6665
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6649..6665
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7042..7385
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      7778..7975
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 taagtcgtaa ttgagtccag atccactgcg gctctgcata tagaataaca aacaaatcgt
       61 atttttcccg aagccaagaa aaaaaggaga ctataacctt tcaagccaga cggaaaacta
      121 tctgcccaag gacaaactgg catgggctgc ttcaatgttc ggctaccttt tttctctttt
      181 aacggggttt ggtatttttt ttctttactt cataaacgaa tgttaaacga ctgtgaaaaa
      241 gccatccctg gattatgcga gagtactctc agggcacccg tccgtttaaa gaccccaggc
      301 caaaataaaa atttcttgaa ggaaaaaaat acacagggcc ttgaactgaa tacaaagcca
      361 tcttttttct cgtttgtcag cacgtcccag aaaaacgcgc agccgtagct cttgataagt
      421 ggcttctgcg ttttgcaacg cttttctgat ctcatgtttt cttatgctgc gggacatcat
      481 tgtcgttgag aaaagtttca atttcaaacg aactgcctac cgttgcattc tccttggaga
      541 agggcctgag tttaggggaa tcacgtctgg ctattacatg ctacatcgtg cagtagctcc
      601 tacatcgcta acaggcaaat cctacaccct ctttaaatcg tttatctttt atatattatc
      661 tggcctaaat ttttacatat aaatatttat catattatga cagctcaaat gaatttccgg
      721 caccaagtca tatgagcact cgctactgtt cgaagaacaa aaaatataaa gcttctacaa
      781 aatttccaga attcacaatg aagctactgt cttctatcga acaagcatgc gatatttgcc
      841 gacttaaaaa gctcaagtgc tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca
      901 actgggagtg tcgctactct cccaaaacca aaaggtctcc gctgactagg gcacatctga
      961 cagaagtgga atcaaggcta gaaagactgg aacagctatt tctactgatt tttcctcgag
     1021 aagaccttga catgattttg aaaatggatt ctttacagga tataaaagca ttgttaacag
     1081 gattatttgt acaagataat gtgaataaag atgccgtcac agatagattg gcttcagtgg
     1141 agactgatat gcctctaaca ttgagacagc atagaataag tgcgacatca tcatcggaag
     1201 agagtagtaa caaaggtcaa agacagttga ctgtatcgat tgactcggca gctcatcatg
     1261 ataactccac aattccgttg gattttatgc ccagggatgc tcttcatgga tttgattggt
     1321 ctgaagagga tgacatgtcg gatggcttgc ccttcctgaa aacggacccc aacaataatg
     1381 ggttctttgg cgacggttct ctcttatgta ttcttcgatc tattggcttt aaaccggaaa
     1441 attacacgaa ctctaacgtt aacaggctcc cgaccatgat tacggataga tacacgttgg
     1501 cttctagatc cacaacatcc cgtttacttc aaagttatct caataatttt cacccctact
     1561 gccctatcgt gcactcaccg acgctaatga tgttgtataa taaccagatt gaaatcgcgt
     1621 cgaaggatca atggcaaatc ctttttaact gcatattagc cattggagcc tggtgtatag
     1681 agggggaatc tactgatata gatgtttttt actatcaaaa tgctaaatct catttgacga
     1741 gcaaggtctt cgagtcaggt tccataattt tggtgacagc cctacatctt ctgtcgcgat
     1801 atacacagtg gaggcagaaa acaaatacta gctataattt tcacagcttt tccataagaa
     1861 tggccatatc attgggcttg aatagggacc tcccctcgtc cttcagtgat agcagcattc
     1921 tggaacaaag acgccgaatt tggtggtctg tctactcttg ggagatccaa ttgtccctgc
     1981 tttatggtcg atccatccag ctttctcaga atacaatctc cttcccttct tctgtcgacg
     2041 atgtgcagcg taccacaaca ggtcccacca tatatcatgg catcattgaa acagcaaggc
     2101 tcttacaagt tttcacaaaa atctatgaac tagacaaaac agtaactgca gaaaaaagtc
     2161 ctatatgtgc aaaaaaatgc ttgatgattt gtaatgagat tgaggaggtt tcgagacagg
     2221 caccaaagtt tttacaaatg gatatttcca ccaccgctct aaccaatttg ttgaaggaac
     2281 acccttggct atcctttaca agattcgaac tgaagtggaa acagttgtct cttatcattt
     2341 atgtattaag agattttttc actaatttta cccagaaaaa gtcacaacta gaacaggatc
     2401 aaaatgatca tcaaagttat gaagttaaac gatgctccat catgttaagc gatgcagcac
     2461 aaagaactgt tatgtctgta agtagctata tggacaatca taatgtcacc ccatattttg
     2521 cctggaattg ttcttattac ttgttcaatg cagtcctagt acccataaag actctactct
     2581 caaactcaaa atcgaatgct gagaataacg agaccgcaca attattacaa caaattaaca
     2641 ctgttctgat gctattaaaa aaactggcca cttttaaaat ccagacttgt gaaaaataca
     2701 ttcaagtact ggaagaggta tgtgcgccgt ttctgttatc acagtgtgca atcccattac
     2761 cgcatatcag ttataacaat agtaatggta gcgccattaa aaatattgtc ggttctgcaa
     2821 ctatcgccca ataccctact cttccggagg aaaatgtcaa caatatcagt gttaaatatg
     2881 tttctcctgg ctcagtaggg ccttcacctg tgccattgaa atcaggagca agtttcagtg
     2941 atctagtcaa gctgttatct aaccgtccac cctctcgtaa ctctccagtg acaataccaa
     3001 gaagcacacc ttcgcatcgc tcagtcacgc cttttctagg gcaacagcaa cagctgcaat
     3061 cattagtgcc actgaccccg tctgctttgt ttggtggcgc caattttaat caaagtggga
     3121 atattgctga tagctcattg tccttcactt tcactaacag tagcaacggt ccgaacctca
     3181 taacaactca aacaaattct caagcgcttt cacaaccaat tgcctcctct aacgttcatg
     3241 ataacttcat gaataatgaa atcacggcta gtaaaattga tgatggtaat aattcaaaac
     3301 cactgtcacc tggttggacg gaccaaactg cgtataacgc gtttggaatc actacaggga
     3361 tgtttaatac cactacaatg gatgatgtat ataactatct attcgatgat gaagataccc
     3421 caccaaaccc aaaaaaagag taaaatgaat cgtagatact gaaaaacccc gcaagttcac
     3481 ttcaactgtg catcgtgcac catctcaatt tctttcattt atacatcgtt ttgccttctt
     3541 ttatgtaact atactcctct aagtttcaat cttggccatg taacctctga tctatagaat
     3601 tttttaaatg actagaatta atgcccatct tttttttgga cctaaattct tcatgaaaat
     3661 atattacgag ggcttattca gaagcttcgc tcatataacg aaaaaaaggg tttggatcga
     3721 acgtaattga gattgattag ttaatactca aaataaaaca gctcctacca ccagtgtaaa
     3781 gtagaacgtt aatagagcca atgtcttcag acagcggccg ctggcctatg gattggtttg
     3841 aagatattgg tgagtagagc acgcagcacg ctgtatttac gtatttaatt ttatatattt
     3901 gtgcatacac tactagggaa gacttgaaaa aaacctagga aatgaaaaaa cgacacagga
     3961 agtcccgtat ttactatttt ttccttcctt ttgatggggc agggcggaaa tagaggatag
     4021 gataagccta ctgcttagct gtttaaacac tggccgtcgt tttacaacgt cgtgactggg
     4081 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4141 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4201 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4261 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     4321 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     4381 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     4441 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4501 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4861 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5161 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5521 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5641 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5761 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5821 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6301 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6361 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6421 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6481 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6541 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6601 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6661 atgaccatga ttacgccagt ttaaactgtg ggagattaga gggcaaagtg tacaaagata
     6721 ccactttgta tacgacgctg tctccatgcg acatgtgtac aggtgccatc atcatgtatg
     6781 gtattccacg ctgtgttgtc ggtgagaacg ttaatttcaa aagtaagggc gattaattaa
     6841 ggatccgctg aagcttcgta cgctgcaggt cgacaaccct taatgttctt cattctacgc
     6901 acggtccttc aattcgcttg gcggagatct atgacaagag ggtcgaactc gcctaagtcg
     6961 taattgagtc agttacgcta gggataacag ggtaatatag ctgtttagct tgcctcgtcc
     7021 ccgccgggtc acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac
     7081 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
     7141 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
     7201 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
     7261 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
     7321 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
     7381 caaccatggg tatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgatccca
     7441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     7501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccgggctc gatcccctcg
     7561 cgagttggtt cagctgctgc ctgaggctgg acgacctcgc ggagttctac cggcagtgca
     7621 aatccgtcgg catccaggaa accagcagcg gctatccgcg catccatgcc cccgaactgc
     7681 aggagtgggg aggcacgatg gccgctttgg tcgacccgga cgggacgctc ctgcgcctga
     7741 tacagaacga attgcttgca ggcatctcat gatcagtact gacaataaaa agattcttgt
     7801 tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat tttaatcaaa
     7861 tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg cgaagttaag
     7921 tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta tactgctgtc
     7981 gattcgatac taacgccgcc atccagtgtc gaaaacgagc tctcgagaac ccttaaagtt
     8041 acgctaggga taacagggta atataggttc ttcattctac gcacggtcct tcaattcgct
     8101 tggcggtcta gaatgacaag agggtcgaac tcgcc
//
