LOCUS       sequence-495634-        8726 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Yeast 2-hybrid vector encoding the human regulator of G protein 
            signaling RGS19 fused to the GAL4 DNA activation domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8726)
  TITLE     Assman lab Y2H vectors
REFERENCE   2  (bases 1 to 8726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pDEST-GADT7-RGS19
FEATURES             Location/Qualifiers
     source          1..8726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1475
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1521..1541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1557..1898
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     1904..1923
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1904..1922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1929..1938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..1967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2004..2028
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2710..2734)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      3154..3341
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3209..3233)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3458..4552)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4553..4958)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5000..5016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5000..5016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5013..5035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5024..5040
                     /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(join(5048..5054,5055..5072,5073..5078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    5169..5202
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5280..5313
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5382..5399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5553..6141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5633..5652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6312..7103,7104..7172))
                     /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     6935..6954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7173..7277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7345..7363
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7401..7423)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7523..7542
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7559..8723
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag
     1201 atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct
     1261 tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc cagttacttg
     1321 aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat
     1381 taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca
     1441 caatatttca agctatacca agcatacaat caactccaag ctttgcaaag atggataaag
     1501 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcca
     1561 attttaatca aagtgggaat attgctgata gctcattgtc cttcactttc actaacagta
     1621 gcaacggtcc gaacctcata acaactcaaa caaattctca agcgctttca caaccaattg
     1681 cctcctctaa cgttcatgat aacttcatga ataatgaaat cacggctagt aaaattgatg
     1741 atggtaataa ttcaaaacca ctgtcacctg gttggacgga ccaaactgcg tataacgcgt
     1801 ttggaatcac tacagggatg tttaatacca ctacaatgga tgatgtatat aactatctat
     1861 tcgatgatga agatacccca ccaaacccaa aaaaagagat ctttaatacg actcactata
     1921 gggcgagcgc cgccatggag tacccatacg acgtaccaga ttacgctcat atggccatgg
     1981 aggccagtga attccaccca tcaacaagtt tgtacaaaaa agcaggctcc gaattcgccc
     2041 ttatgcccac cccgcatgag gctgagaagc agatcacagg gccagaggag gcggaccggc
     2101 ccccttcaat gtccagtcat gatacagcct ctccagcggc ccccagccgc aacccctgct
     2161 gcctgtgctg gtgctgctgc tgtagctgct cctggaacca agagcggcgg cgcgcgtggc
     2221 aggcctcccg ggagagcaag ctgcagcccc tccccagctg tgaagtatgt gccacgccaa
     2281 gtcctgagga ggtgcagagc tgggcgcagt cttttgacaa gctgatgcac agcccagcgg
     2341 gacgcagcgt gttccgggcg ttcctgcgga cagagtacag cgaggagaac atgctcttct
     2401 ggttggcctg cgaggagctg aaggccgagg ccaaccagca tgtggtagac gagaaggcga
     2461 ggctcatcta cgaggactac gtatccatcc tgtcccccaa ggaggtgagc ctggactccc
     2521 gtgtgcggga gggcatcaac aagaagatgc aggagccgtc cgcacacacg ttcgacgacg
     2581 cgcagctgca gatctacacg ctcatgcacc gggactccta cccccgcttc ctcagctctc
     2641 ccacctaccg tgccctgctg ctgcaggggc catcacagtc ctcctccgag gcctagaagg
     2701 gcgaattcga cccagctttc ttgtacaaag tggttgatgg gtgggcatcg atacgggatc
     2761 catcgagctc gagctgcaga tgaatcgtag atactgaaaa accccgcaag ttcacttcaa
     2821 ctgtgcatcg tgcaccatct caatttcttt catttataca tcgttttgcc ttcttttatg
     2881 taactatact cctctaagtt tcaatcttgg ccatgtaacc tctgatctat agaatttttt
     2941 aaatgactag aattaatgcc catctttttt ttggacctaa attcttcatg aaaatatatt
     3001 acgagggctt attcagaagc tttggacttc ttcgccagag gtttggtcaa gtctccaatc
     3061 aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa gggtcaaatc
     3121 gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt tatgattttt
     3181 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     3241 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     3301 gtatagcatg aggtcgctct tattgaccac acctctaccg gccggtcgaa attcccctac
     3361 cctatgaaca tattccattt tgtaatttcg tgtcgtttct attatgaatt tcatttataa
     3421 agtttatgta caaatatcat aaaaaaagag aatcttttta agcaaggatt ttcttaactt
     3481 cttcggcgac agcatcaccg acttcggtgg tactgttgga accacctaaa tcaccagttc
     3541 tgatacctgc atccaaaacc tttttaactg catcttcaat ggccttacct tcttcaggca
     3601 agttcaatga caatttcaac atcattgcag cagacaagat agtggcgata gggttgacct
     3661 tattctttgg caaatctgga gcagaaccgt ggcatggttc gtacaaacca aatgcggtgt
     3721 tcttgtctgg caaagaggcc aaggacgcag atggcaacaa acccaaggaa cctgggataa
     3781 cggaggcttc atcggagatg atatcaccaa acatgttgct ggtgattata ataccattta
     3841 ggtgggttgg gttcttaact aggatcatgg cggcagaatc aatcaattga tgttgaacct
     3901 tcaatgtagg aaattcgttc ttgatggttt cctccacagt ttttctccat aatcttgaag
     3961 aggccaaaac attagcttta tccaaggacc aaataggcaa tggtggctca tgttgtaggg
     4021 ccatgaaagc ggccattctt gtgattcttt gcacttctgg aacggtgtat tgttcactat
     4081 cccaagcgac accatcacca tcgtcttcct ttctcttacc aaagtaaata cctcccacta
     4141 attctctgac aacaacgaag tcagtacctt tagcaaattg tggcttgatt ggagataagt
     4201 ctaaaagaga gtcggatgca aagttacatg gtcttaagtt ggcgtacaat tgaagttctt
     4261 tacggatttt tagtaaacct tgttcaggtc taacactacc tgtaccccat ttaggaccac
     4321 ccacagcacc taacaaaacg gcatcagcct tcttggaggc ttccagcgcc tcatctggaa
     4381 gtgggacacc tgtagcttcg atagcagcac caccaattaa atgattttcg aaatcgaact
     4441 tgacattgga acgaacatca gaaatagctt taagaacctt aatggcttcg gctgtgattt
     4501 cttgaccaac gtggtcacct ggcaaaacga cgatcttctt aggggcagac attagaatgg
     4561 tatatccttg aaatatatat atatattgct gaaatgtaaa aggtaagaaa agttagaaag
     4621 taagacgatt gctaaccacc tattggaaaa aacaataggt ccttaaataa tattgtcaac
     4681 ttcaagtatt gtgatgcaag catttagtca tgaacgcttc tctattctat atgaaaagcc
     4741 ggttccggcg ctctcacctt tcctttttct cccaattttt cagttgaaaa aggtatatgc
     4801 gtcaggcgac ctctgaaatt aacaaaaaat ttccagtcat cgaatttgat tctgtgcgat
     4861 agcgcccctg tgtgttctcg ttatgttgag gaaaaaaata atggttgcta agagattcga
     4921 actcttgcat cttacgatac ctgagtattc ccacagttgg gggatctcga ctctagctag
     4981 aggatcaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5161 gccagctgat aacttcgtat aatgtatgct atacgaagtt attaggtctg aagaggagtt
     5221 tacgtccagc caagctagct tggctgcagg tcgagcggcc gcgatccgga acccttaata
     5281 taacttcgta taatgtatgc tatacgaagt tatcagctgc attaatgaat cggccaacgc
     5341 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5401 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5461 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5521 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5581 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5641 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5701 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5761 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5821 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5881 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5941 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6001 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6061 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6121 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6181 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6241 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6301 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6361 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6421 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6481 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6541 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6601 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6661 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6721 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6781 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6841 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6901 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6961 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7021 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7081 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7141 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7201 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7261 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     7321 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     7381 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     7441 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     7501 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccataac
     7561 gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata tacaggcaac
     7621 acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc gttgcatttt
     7681 cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc tattctctag
     7741 ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg aagacgcact
     7801 ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc gaataccgct
     7861 tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat atccctatat
     7921 aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct ctacattttt
     7981 tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac tattcataga
     8041 gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat agagacaaaa
     8101 tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta tcactttctg
     8161 ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc ctttatcttg
     8221 aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga gtcaggcttt
     8281 ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac ggacctacag
     8341 tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa aagtaatcta
     8401 agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca aaaaagaagt
     8461 atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg taaaaatgca
     8521 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ttacaaaaat
     8581 gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg ttctgtaaaa
     8641 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttctaca
     8701 aaatgaagca cagatgcttc gttgct
//
