LOCUS       sequence-154426-        7407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast expression vector p414GPD modified such that it is Gateway 
            converted and introduces a 3xFLAG tag at the N-terminus of the 
            insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7407)
  AUTHORS   McCann KL, Charette JM, Vincent NG, Baserga SJ
  TITLE     A protein interaction map of the LSU processome.
  JOURNAL   Genes Dev. 2015 Apr 15;29(8):862-75. doi: 10.1101/gad.256370.114. 
            Epub 2015 Apr 15.
  PUBMED    25877921
REFERENCE   2  (bases 1 to 7407)
  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: 
            "10.1101/gad.256370.114"; journalName: "Genes Dev"; date: 
            "2015-04-15- 15"; volume: "29"; issue: "8"; pages: "862-75"
COMMENT     Sequence Label: p414GPD-3xFLAG-GW
FEATURES             Location/Qualifiers
     source          1..7407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2803..3445
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3338..3361
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3451..3467
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3451..3467
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3484..3549
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    3550..3674
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3699..3704,3705..3722,3723..3729)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3783..4442
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3849..3868)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4784..5089
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4968..4987
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5130..5254)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      5257..5504
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5273..5291)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(5522..5541)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5523..5541)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5551..5568)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5551..5567)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5560..5582)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5712..6167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5799..5818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6009..6030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6266..6940)
                     /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(6941..7221)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(7238..7257)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7357..7379
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctctcgagtt tatcattatc aatactgcca
     2821 tttcaaagaa tacgtaaata attaatagta gtgattttcc taactttatt tagtcaaaaa
     2881 attagccttt taattctgct gtaacccgta catgccaaaa tagggggcgg gttacacaga
     2941 atatataaca ctgatggtgc ttgggtgaac aggtttattc ctggcatcca ctaaatataa
     3001 tggagcccgc tttttaagct ggcatccaga aaaaaaaaga atcccagcac caaaatattg
     3061 ttttcttcac caaccatcag ttcataggtc cattctctta gcgcaactac agagaacagg
     3121 gcacaaacag gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa
     3181 atgatgacac aaggcaattg acccacgcat gtatctatct cattttctta caccttctat
     3241 taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa aggtttaaac cagttccctg
     3301 aaattattcc cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct
     3361 gtaaatctat ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt
     3421 taaaacacca agaacttagt ttcgacggat tctagaacta gtggatcccc cgggctgcag
     3481 atggactaca aagaccatga cggtgattat aaagatcatg atatcgatta caaggatgac
     3541 gatgacaaga caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     3601 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     3661 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     3721 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     3781 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3841 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     3901 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3961 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     4021 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     4081 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     4141 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     4201 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     4261 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     4321 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     4381 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     4441 aaagatctgg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     4501 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     4561 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     4621 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     4681 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     4741 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     4801 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     4861 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     4921 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     4981 cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc tcagccaccg
     5041 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     5101 ccttatacac agccagtctg caggtcgacc atagtgactg gatatgttgt gttttacagt
     5161 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     5221 acgtttctcg ttcagctttc ttgtacaaag tggtagtcat gtaattagtt atgtcacgct
     5281 tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg
     5341 aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat
     5401 ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa
     5461 ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc
     5521 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5581 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5641 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5701 cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5761 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5821 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5881 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5941 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6001 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6061 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6121 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat
     6181 tttctcctta cgcatctgtg cggtatttca caccgcatag gcaagtgcac aaacaatact
     6241 taaataaata ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt
     6301 tgttagagtc ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc
     6361 catttaatct aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat
     6421 gtaagctttc ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa
     6481 gttcacctgt cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg
     6541 aagctgcact gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca
     6601 aaccgaggaa ctcttggtat tcttgccacg actcatctcc atgcagttgg acgatatcaa
     6661 tgccgtaatc attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa
     6721 ccaagtattt cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc
     6781 ttgcaataac cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt
     6841 cagcatcgga atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca
     6901 ccaatggacc agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt
     6961 gcttaatcac gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc
     7021 cttttctttt ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa
     7081 gattgtacgt aaggtgacaa gctatttttc aataaagaat atcttccact actgccatct
     7141 ggcgtcataa ctgcaaagta cacatatatt acgatgctgt ctattaaatg cttcctatat
     7201 tatatatata gtaatgtcgt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     7261 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     7321 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     7381 tttcaccgtc atcaccgaaa cgcgcga
//
