LOCUS       sequence-254059-        8463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8463)
  AUTHORS   Grossmann A, Benlasfer N, Birth P, Hegele A, Wachsmuth F, Apelt L, 
            Stelzl U
  TITLE     Phospho-tyrosine dependent protein-protein interaction network.
  JOURNAL   Mol Syst Biol. 2015 Mar 26;11(3):794.
  PUBMED    25814554
REFERENCE   2  (bases 1 to 8463)
  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: "Mol Syst 
            Biol."; date: "2015-03-26"; volume: "11(3)"; pages: "794"
COMMENT     Sequence Label: pASZ-C-DM
FEATURES             Location/Qualifiers
     source          1..8463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(451..1039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(531..550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1210..2001,2002..2070))
                     /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     1833..1852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2071..2175)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2243..2261
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2299..2321)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2421..2440
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2649..2665
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2937..3061
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3235..3337
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3338..3997
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3404..3423)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4339..4644
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTADMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(4685..4809)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4826..5073
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4842..4860)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5091..5110)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5092..5110)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5120..5136)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5364..5383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5424..5440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5424..5440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5437..5459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5448..5464
                     /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(5472..5478,5479..5496,5497..5502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5517..5538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5604..5979
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             5980..7695
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     primer_bind     complement(7915..7932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cttctgcaca gaagtgtgta tatatatatt tatatatagt cgatgtgcaa aagcctacat
       61 tatttttctt ttgattttta tttttttgtt caagtaggtt catgccgaca gtaaatcctt
      121 atgccctacc taaatataaa cacccaaata tatatatata tatatatata tataaataat
      181 aaaaacggaa aggaaataga ataataataa taatactaca cgtttacgta tgtaactcat
      241 tctattattc agactttgat attaatagaa ttattactac taggtttcct cacttcaaaa
      301 tctataaata taccgtcatg ctctaatcga tgctcactca aaggcggtaa tacggttatc
      361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      961 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1021 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1081 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1141 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1201 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1261 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1321 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     1381 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1441 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1501 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1561 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1621 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1681 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1741 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1801 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1861 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1921 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1981 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2041 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     2101 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2161 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2221 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     2281 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2341 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2401 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     2461 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     2521 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2581 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2641 acgacgttgt aaaacgacgg ccagtgaatt cgagctcctt ttgctggcat ttcttctaga
     2701 agcaaaaaga gcgatgcgtc ttttccgctg aaccgttcca gcaaaaaaga ctaccaacgc
     2761 aatatggatt gtcagaatca tataaaagag aagcaaataa ctccttgtct tgtatcaatt
     2821 gcattataat atcttcttgt tagtgcaata tcatatagaa gtcatcgaaa tagatattaa
     2881 gaaaaacaaa ctgtacaatc aatcaatcat cacataaaat ggggtcgacg gatctcacaa
     2941 gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     3001 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     3061 ggcggccgcg ggtgatgctg ccaacttagc ggccgctaag ttggcagcat cacccgacgc
     3121 actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg
     3181 tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg
     3241 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3301 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     3361 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3421 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     3481 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     3541 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     3601 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     3661 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3721 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3781 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3841 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     3901 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     3961 acaacagtac tgcgatgagt ggcagggcgg ggcgtaatct agaggatccg gcttactaaa
     4021 agccagataa cagtatgcgt atttgcgcgc tgattttcgc ggtataagaa tatatactga
     4081 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     4141 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     4201 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     4261 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     4321 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     4381 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     4441 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
     4501 catatcgggg atgaaagctg gcgcatgatg accgccgata tggccagtgt gccggtctcc
     4561 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     4621 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     4681 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     4741 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     4801 caaagtggta agtaactaac tcgagtcatg taattagtta tgtcacgctt acattcacgc
     4861 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     4921 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     4981 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     5041 aaggttttgg gacgctcgaa ggctttaatt tgcggccggt acccaattcg ccctatagtg
     5101 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     5161 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     5221 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     5281 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5341 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5401 cgcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5461 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5521 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5581 tcgtgccagc agatctgaat taattcttga ataatacata acttttctta aaagaatcaa
     5641 agacagataa aatttaagag atattaaata ttagtgagaa gccgagaatt ttgtaacacc
     5701 aacataacac tgacatcttt aacaactttt aattatgata catttcttac gtcatgattg
     5761 attattacag ctatgctgac aaatgactct tgttgcatgg ctacgaaccg ggtaatacta
     5821 agtgattgac tcttgctgac cttttattaa gaactaaatg gacaatatta tggagcattt
     5881 catgtataaa ttggtgcgta aaatcgttgg atctctcttc taagtacatc ctactataac
     5941 aatcaagaaa aacaagaaaa tcggacaaaa caatcaagta tggattctag aacagttggt
     6001 atattaggag ggggacaatt gggacgtatg attgttgagg cagcaaacag gctcaacatt
     6061 aagacggtaa tactagatgc tgaaaattct cctgccaaac aaataagcaa ctccaatgac
     6121 cacgttaatg gctccttttc caatcctctt gatatcgaaa aactagctga aaaatgtgat
     6181 gtgctaacga ttgagattga gcatgttgat gttcctacac taaagaatct tcaagtaaaa
     6241 catcccaaat taaaaattta cccttctcca gaaacaatca gattgataca agacaaatat
     6301 attcaaaaag agcatttaat caaaaatggt atagcagtta cccaaagtgt tcctgtggaa
     6361 caagccagtg agacgtccct attgaatgtt ggaagagatt tgggttttcc attcgtcttg
     6421 aagtcgagga ctttggcata cgatggaaga ggtaacttcg ttgtaaagaa taaggaaatg
     6481 attccggaag ctttggaagt actgaaggat cgtcctttgt acgccgaaaa atgggcacca
     6541 tttactaaag aattagcagt catgattgtg aggtctgtta acggtttagt gttttcttac
     6601 ccaattgtag agactatcca caaggacaat atttgtgact tatgttatgc gcctgctaga
     6661 gttccggact ccgttcaact taaggcgaag ttgttggcag aaaatgcaat caaatctttt
     6721 cccggttgtg gtatatttgg tgtggaaatg ttctatttag aaacagggga attgcttatt
     6781 aacgaaattg ccccaaggcc tcacaactct ggacattata ccattgatgc ttgcgtcact
     6841 tctcaatttg aagctcattt gagatcaata ttggatttgc caatgccaaa gaatttcaca
     6901 tctttctcca ccattacaac gaacgccatt atgctaaatg ttcttggaga caaacataca
     6961 aaagataaag agctagaaac ttgcgaaaga gcattggcga ctccaggttc ctcagtgtac
     7021 ttatatggaa aagagtctag acctaacaga aaagtaggtc acataaatat tattgcctcc
     7081 agtatggcgg aatgtgaaca aaggctgaac tacattacag gtagaactga tattccaatc
     7141 aaaatctctg tcgctcaaaa gttggacttg gaagcaatgg tcaaaccatt ggttggaatc
     7201 atcatgggat cagactctga cttgccggta atgtctgccg catgtgcggt tttaaaagat
     7261 tttggcgttc catttgaagt gacaatagtc tctgctcata gaactccaca taggatgtca
     7321 gcatatgcta tttccgcaag caagcgtgga attaaaacaa ttatcgctgg agctggtggg
     7381 gctgctcact tgccaggtat ggtggctgca atgacaccac ttcctgtcat cggtgtgccc
     7441 gtaaaaggtt cttgtctaga tggagtagat tctttacatt caattgtgca aatgcctaga
     7501 ggtgttccag tagctaccgt cgctattaat aatagtacga acgctgcgct gttggctgtc
     7561 agactgcttg gcgcttatga ttcaagttat acaacgaaaa tggaacagtt tttattaaag
     7621 caagaagaag aagttcttgt caaagcacaa aagttagaaa ctgtcggtta cgaagcttat
     7681 ctagaaaaca agtaatatat aagtttattg atatacttgt acagcaaata attataaaat
     7741 gatataccta ttttttaggc tttgttatga ttacatcaaa tgtggacttc atacatagaa
     7801 atcaacgctt acaggtgtcc ttttttaaga atttcataca taagatctgc tgcattaatg
     7861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagcat cgatgctaga
     7981 gtcgaatgga aattacgtac tttcaacgca agagcaagac aaaagaaagc ctaaaactgt
     8041 cttttcgtgt cggtcgtcca atatcatcgt aaacgtgtgt agagccttaa attctaattt
     8101 ctaactttaa attttttttg ctattataat actaatttct aactttcaac ctattttaca
     8161 tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt
     8221 tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagaa
     8281 atatagtctt caaggttata ttatacaggt ttatatatta ttttacttat ttttgttgtt
     8341 ctttttcttg gagtatttag cattaacaac ttcgacccat cgcgatgacg aaattgcacg
     8401 tttaaaggag aagtcggaat cgttgcaaga agaaataaat tagtttgacg ttttattcct
     8461 ttt
//
