LOCUS       sequence-386973-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of GAL4 DNA-binding domain (BD)-tepsin fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Mattera R, Guardia CM, Sidhu SS, Bonifacino JS
  TITLE     Bivalent Motif-Ear Interactions Mediate the Association of the 
            Accessory Protein Tepsin with the AP-4 Adaptor Complex.
  JOURNAL   J Biol Chem. 2015 Dec 25;290(52):30736-49. doi: 
            10.1074/jbc.M115.683409. Epub 2015 Nov 5.
  PUBMED    26542808
REFERENCE   2  (bases 1 to 7091)
  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"; doi: 
            "10.1074/jbc.M115.683409"; journalName: "J Biol Chem"; date: 
            "2015-12-25- 25"; volume: "290"; issue: "52"; pages: "30736-49"
COMMENT     Sequence Label: pGBT9-tepsin
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             434..874
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     terminator      2489..2676
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2544..2568)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2729..3403)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3404..3505)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3519..3535)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3519..3535)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3532..3554)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3543..3559
                     /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(3567..3573,3574..3591,3592..3597))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3612..3633
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3750..3767)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3921..4509)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4001..4020)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4680..5471,5472..5540))
                     /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     5303..5322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5541..5645)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5713..5731
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5769..5791)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5891..5910
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      5927..7091
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac
      121 agcaccaaca gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact
      181 ttttccttcc ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt
      241 ccagttactt gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcatggctg ccgcgccgcc
      901 gctacgggac cgcctgagct ttctacaccg gctcccgatt ctcctgaagg ggacgtccga
      961 tgatgatgtc ccgtgtccgg gctacctgtt tgaagagatt gctaaaatct cccacgagtc
     1021 tccgggcagc agccagtgcc tgctggagta cctcctgagc cgcctgcaca gcagctccgg
     1081 ccacgggaag ctcaaggtgc tgaagatcct gctctatctg tgcagccacg gctcctcctt
     1141 cttcctgctc atcctcaaac gcaactctgc cttcatccag gaagctgcag cttttgcagg
     1201 gcccccagat cctctgcacg ggaacagctt gtaccagaag gttcgcgcgg ccgcgcagga
     1261 cttggggagc accctgttct cggacaccgt gttgccgctg gctccctccc agcctctggg
     1321 gaccccgcct gccacaggca tgggctccca ggccaggccg cacagcaccc tccagggttt
     1381 cggctacagc aaggaacacg gccgcacggc tgtgaggcat cagcctgggc aggccggagg
     1441 gggctgggat gagctggaca gcggccccag ctctcagaac tcctcccaga acagtgacct
     1501 gagcagggtc tcggactcgg gcagtcattc cggcagcgac agccattcag gggccagccg
     1561 ggagccgggt gacctggcag aaagggtcga ggtggtggcc ctgagtgact gtcagcagga
     1621 gttgagcttg gtgaggactg tgactcgggg accacgcgcc ttcctgagcc gcgaggaggc
     1681 acagcacttc atcaaagcgt gtggactgct caactgtgag gccgtgctgc agctgctgac
     1741 ctgccacctg cgtgggacca gtgaatgcac gcagctgagg gcgctgtgtg ccatcgcctc
     1801 cctggggagc agcgacctcc tcccccagga gcacatcctc ctccgcaccc ggccgtggct
     1861 gcaggagctc agcatgggca gcccgggacc tgtgaccaac aaggccacca agatcctgag
     1921 gcactttgag gcctcctgtg ggcagctgtc ccctgcccgg ggcacctcag ctgagcctgg
     1981 ccccacagcc gccctcccag gcccatctga cctgctgacc gacgctgtgc ctctccctgg
     2041 gagccaggtc ttcctgcagc ctctgagttc aaccccggtc tcgtcccgga gccctgctcc
     2101 ctcatctggg atgccgtcca gccctgtgcc caccccaccc ccagatgcct cccccattcc
     2161 agcccccgga gaccccagcg aggccgaggc cagactggca gaaagcaggc ggtggagacc
     2221 tgaacggatc ccagggggca cggacagccc aaagagaggc cccagcagct gtgcgtggag
     2281 ccgcgactcc ttgtttgctg gcatggagct ggtggcctgt ccccgcctgg tgggggctgg
     2341 ggctgctgcg ggagagtcct gtcctgatgc tccccgcgcc ccccaaacat cgtcccagag
     2401 gacagcagcc aaagagcctc ctggctcaga gccgtcagct ttcgcgttcc tgaacgcctg
     2461 actcgacctg cagccaagct aattccgggc gaatttctta tgatttatga tttttattat
     2521 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     2581 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     2641 gcatgaggtc gctcttattg accacacctc taccggcatg ccggcaagtg cacaaacaat
     2701 acttaaataa atactactca gtaataacct atttcttagc atttttgacg aaatttgcta
     2761 ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa
     2821 cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa
     2881 aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca
     2941 aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct
     3001 gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg
     3061 gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat
     3121 caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc
     3181 caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt
     3241 tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata atacccagca
     3301 agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt
     3361 tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg
     3421 tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct
     3481 ctccttttct tttttcgacc gaattaattc gtaatcatgt catagctgtt tcctgtgtga
     3541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4081 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4321 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4441 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4621 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4741 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4861 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5101 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5341 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5641 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5701 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     5761 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5821 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5881 cttaactatg cggcatcaga gcagattgta ctgagagtgc accataacgc atttaagcat
     5941 aaacacgcac tatgccgttc ttctcatgta tatatatata caggcaacac gcagatatag
     6001 gtgcgacgtg aacagtgagc tgtatgtgcg cagctcgcgt tgcattttcg gaagcgctcg
     6061 ttttcggaaa cgctttgaag ttcctattcc gaagttccta ttctctagct agaaagtata
     6121 ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa gacgcacttt caaaaaacca
     6181 aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc cacaaacatt
     6241 gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa cctacccatc
     6301 cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta tgtttatctc
     6361 tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt gaatcgaaaa
     6421 caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata gaagaaaccg
     6481 ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt cacaaagtat
     6541 gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa aaaatgcacc
     6601 cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt ttatggaaga
     6661 gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg caaaaagtta
     6721 tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag atgctttgtt
     6781 agaaaaatag cgctctcggg atgcattttt gtagaacaaa aaagaagtat agattctttg
     6841 ttggtaaaat agcgctctcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     6901 tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt acaaaaatga agcacagatt
     6961 cttcgttggt aaaatagcgc tttcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     7021 ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttctacaaa atgaagcaca
     7081 gatgcttcgt t
//
