LOCUS       Exported                3858 bp ds-DNA     circular SYN 06-APR-2022
DEFINITION  vertebrate expression of full-length temperature-sensitive Gal80 
            with stop.
ACCESSION   .
VERSION     .
KEYWORDS    pME-Gal80ts
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3858)
  AUTHORS   Fujimoto E, Gaynes B, Brimley CJ, Chien CB, Bonkowsky JL
  TITLE     Gal80 intersectional regulation of cell-type specific expression in 
            vertebrates.
  JOURNAL   Dev Dyn. 2011 Oct;240(10):2324-34. doi: 10.1002/dvdy.22734. Epub 
            2011 Sep 8.
  PUBMED    21905164
REFERENCE   2  (bases 1 to 3858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 6, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3858
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             106..1413
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae gal80 mutant allele"
                     /allele="temperature-sensitive allele encoding G203V and 
                     G323R mutations"
                     /product="mutant transcriptional repressor that inhibits 
                     GAL4, but only at temperatures below 29(o)C"
                     /label=GAL80ts
                     /note="GAL80ts binds to and inhibits the transcriptional 
                     activation domain of GAL4."
                     /translation="MDYNKRSSVSTVPNAAPIRVGFVGLNAAKGWAIKTHYPAILQLSS
                     QFQITALYSPKIETSIATIQRLKLSNATAFPTLESFASSSTIDMIVIAIQVASHYDVVM
                     PLLEFSKNNPNLKYLFVEWALACSLDQAESIYKAAAERGVQTIISLQGRKSPYILRAKE
                     LISQGYIGDINSIEIAGNGGWYGYERPVKSPKYIYEIGNVVDLVTTTFGHTIDILQYMT
                     SSYFSRINAMVFNNIPEQELIDERGNRLGQRVPKTVPDHLLFQGTLLNGNVPVSCSFKG
                     GKPTKKFTKNLVIDIHGTKGDLKLEGDAGFAEISNLVLYYSRTRANDFPLANGQQAPLD
                     PGYDAGKEIMEVYHLRNYNAIVGNIHRLYQSISDFHFNTKKIPELPSQFVMQGFDFEGF
                     PTLMDALILHRLIESVYKSNMMGSTLNVSNISHYSL"
     protein_bind    complement(1415..1514)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1531..1550)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1532..1550)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1555..1571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1555..1571)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1673..1692)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1684..2493
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1751..1770)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2640..3228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3129..3148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3382..3399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3558..3585
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      3677..3763
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3812..3834
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3826..3843
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3827..3843
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ctaccatgga ctacaacaag
      121 agatcttcgg tctcaaccgt gcctaatgca gctcccataa gagtcggatt cgtcggtctc
      181 aacgcagcca aaggatgggc aatcaagaca cattaccccg ccatactgca actatcgtca
      241 caatttcaaa tcactgcctt atacagtcca aaaattgaga cttctattgc caccatccag
      301 cgtctaaaat tgagtaatgc cactgctttt cccactttag agtcatttgc atcatcttcc
      361 actatagata tgatagtgat agctatccaa gtggccagtc attatgacgt tgttatgcct
      421 ctcttggaat tctccaaaaa taatccgaac ctcaagtatc ttttcgtaga atgggccctt
      481 gcatgttcac tagatcaagc cgaatccatt tataaggctg ctgctgaacg tggggttcaa
      541 accatcatct ctttacaagg tcgtaaatca ccatatattt tgagagcaaa agaattaata
      601 tctcaaggct atatcggcga cattaattcg atcgagattg ctggaaatgg cggttggtac
      661 ggctacgaaa ggcctgttaa atcaccaaaa tacatctatg aaatcgggaa cgttgtagat
      721 ctggtaacca caacatttgg tcacacaatc gatattttac aatacatgac aagttcgtac
      781 ttttccagga taaatgcaat ggttttcaat aatattccag agcaagagct gatagatgag
      841 cgtggtaacc gattgggcca gcgagtccca aagacagtac cggatcatct tttattccaa
      901 ggcacattgt taaatggcaa tgttccagtg tcatgcagtt tcaaaggtgg caaacctacc
      961 aaaaaattta ccaaaaattt ggtcattgat attcacggta ccaagggaga tttgaaactt
     1021 gaaggcgatg ccggattcgc agaaatttca aatctggtcc tttactacag tagaactaga
     1081 gcaaacgact tcccgctagc caatggacaa caagctcctt tagacccggg gtatgatgca
     1141 ggtaaagaaa tcatggaagt atatcattta cgaaattata atgccattgt gggtaacatt
     1201 catcgactgt atcaatctat ctctgacttc cacttcaata caaagaaaat tcctgaatta
     1261 ccctcacaat ttgtaatgca aggtttcgat ttcgaaggct ttcccacctt gatggatgct
     1321 ctgatattac acaggttaat cgagagcgtt tataaaagta acatgatggg ctccacatta
     1381 aacgttagca atatctcgca ttatagttta taatacccag ctttcttgta caaagttggc
     1441 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1501 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1561 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1621 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1681 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1741 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1801 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1861 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1921 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1981 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2041 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2101 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2161 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2221 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2281 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2341 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2401 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2461 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2521 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2581 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2761 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2821 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2881 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2941 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3001 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3061 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3121 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3181 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3241 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3301 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3361 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3421 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3481 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3541 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3601 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3661 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3721 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3781 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3841 agtcttaagc tcgggccc
//
