LOCUS       sequence-329054-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An empty HIS3 plasmid for translation factor eIF4E complementation 
            test in yeast, strain Jo55. Golden Gate cloning using BsaI. 
            Selection with RFP after cloning. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  AUTHORS   Lebedeva MV, Nikonova EY, Terentiev AA, Taranov VV, Babakov AV, 
            Nikonov OS
  TITLE     VPg of Potato Virus Y and Potato Cap-Binding eIF4E Factors: 
            Selective Interaction and Its Supposed Mechanism.
  JOURNAL   Biochemistry (Mosc). 2021 Sep;86(9):1128-1138. doi: 
            10.1134/S000629792109008X.
  PUBMED    34565316
REFERENCE   2  (bases 1 to 7568)
  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"; journalName: 
            "Biochemistry (Mosc)"; date: "2021-09"; volume: "86"; issue: "9"; 
            pages: "1128-1138"
COMMENT     Sequence Label: pJo55
FEATURES             Location/Qualifiers
     source          1..7568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..644
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     538..561
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     650..666
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     650..666
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    781..802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(817..822,823..840,841..847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    855..871
                     /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)."
     RBS             889..900
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             907..1584
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1654..1901
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1670..1688)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1936..1955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1937..1955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1965..1982)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1965..1981)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1974..1996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2120..2575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2207..2226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2417..2438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2835..3497)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3498..3685)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(3832..3851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3951..3969)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      4010..5352
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    4342..4389
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        5379..5483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5484..5552,5553..6344)
                     /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(5702..5721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6515..7103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7004..7023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7257..7274
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7391..7412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7427..7432,7433..7450,7451..7457)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7465..7481
                     /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     7489..7505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7489..7505
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7524..7544
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7526..7544
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tcattatcaa tactcgccat ttcaaagaat acgtaaataa ttaatagtag tgattttcct
       61 aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac atgcccaaaa
      121 tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac agtttattcc
      181 tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa aaaaaaagaa
      241 tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc attctcttag
      301 cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct caatggagtg
      361 atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat gtatctatct
      421 cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag ctgaaaaaaa
      481 aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat ataaagacgg
      541 taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc tacttttata
      601 gttagtcttt tttttagttt taaaacacca gaacttagtt tcgacggatt ctagaactag
      661 tggatcccca aaatgagacc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      721 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      781 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      841 tatgttgtgt ggaattgtga gcggataaca atttcacaca tactagagaa agaggagaaa
      901 tactagatgg cttcctccga agacgttatc aaagagttca tgcgtttcaa agttcgtatg
      961 gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac
     1021 gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg
     1081 gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca cccggctgac
     1141 atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg tgttatgaac
     1201 ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga cggtgagttc
     1261 atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt tatgcagaaa
     1321 aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa
     1381 ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc tgaagttaaa
     1441 accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac cgacatcaaa
     1501 ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga acgtgctgaa
     1561 ggtcgtcact ccaccggtgc ttaataacgc tgatagtgct agtgtagatc gctactagag
     1621 ccaggcggtc tctactgggg gctgcaggaa ttctcatgta attagttatg tcacgcttac
     1681 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     1741 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     1801 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     1861 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cgtcgacctc gagggggggc
     1921 ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     1981 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2041 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg cgcccttccc aacagttgcg
     2101 cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2161 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2341 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2401 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2461 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2521 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga
     2581 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagat ccgtcgagtt
     2641 caagagaaaa aaaaagaaaa agcaaaaaga aaaaaggaaa gcgcgcctcg ttcagaatga
     2701 cacgtataga atgatgcatt accttgtcat cttcagtatc atactgttcg tatacatact
     2761 tactgacatt cataggtata catatataca catgtatata tatcgtatgc tgcagcttta
     2821 aataatcggt gtcactacat aagaacacct ttggtggagg gaacatcgtt ggtaccattg
     2881 ggcgaggtgg cttctcttat ggcaaccgca agagccttga acgcactctc actacggtga
     2941 tgatcattct tgcctcgcag acaatcaacg tggagggtaa ttctgctagc ctctgcaaag
     3001 ctttcaagaa aatgcgggat catctcgcaa gagagatctc ctactttctc cctttgcaaa
     3061 ccaagttcga caactgcgta cggcctgttc gaaagatcta ccaccgctct ggaaagtgcc
     3121 tcatccaaag gcgcaaatcc tgatccaaac ctttttactc cacgcacggc ccctagggcc
     3181 tctttaaaag cttgaccgag agcaatcccg cagtcttcag tggtgtgatg gtcgtctatg
     3241 tgtaagtcac caatgcactc aacgattagc gaccagccgg aatgcttggc cagagcatgt
     3301 atcatatggt ccagaaaccc tatacctgtg tggacgttaa tcacttgcga ttgtgtggcc
     3361 tgttctgcta ctgcttctgc ctctttttct gggaagatcg agtgctctat cgctagggga
     3421 ccacccttta aagagatcgc aatctgaatc ttggtttcat ttgtaatacg ctttactagg
     3481 gctttctgct ctgtcatctt tgccttcgtt tatcttgcct gctcattttt tagtatattc
     3541 ttcgaagaaa tcacattact ttatataatg tataattcat tatgtgataa tgccaatcgc
     3601 taagaaaaaa aaagagtcat ccgctaggtg gaaaaaaaaa aatgaaaatc attaccgagg
     3661 cataaaaaaa tatagagtgt actagaggag gccaagagta atagaaaaag aaaattgcgg
     3721 gaaaggactg tgttatgact tccctgacta atgccgtgtt caaacgatac ctggcagtga
     3781 ctcctagcgc tcaccaagct cttaaaacgg gaatttatgg tgcactctca gtacaatctg
     3841 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     3901 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacaaagg gcctcgtgat
     3961 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagtatg atccaatatc
     4021 aaaggaaatg atagcattga aggatgagac taatccaatt gaggagtggc agcatataga
     4081 acagctaaag ggtagtgctg aaggaagcat acgatacccc gcatggaatg ggataatatc
     4141 acaggaggta ctagactacc tttcatccta cataaataga cgcatataag tacgcattta
     4201 agcataaaca cgcactatgc cgttcttctc atgtatatat atatacaggc aacacgcaga
     4261 tataggtgcg acgtgaacag tgagctgtat gtgcgcagct cgcgttgcat tttcggaagc
     4321 gctcgttttc ggaaacgctt tgaagttcct attccgaagt tcctattctc tagaaagtat
     4381 aggaacttca gagcgctttt gaaaaccaaa agcgctctga agacgcactt tcaaaaaacc
     4441 aaaaacgcac cggactgtaa cgagctacta aaatattgcg aataccgctt ccacaaacat
     4501 tgctcaaaag tatctctttg ctatatatct ctgtgctata tccctatata acctacccat
     4561 ccacctttcg ctccttgaac ttgcatctaa actcgacctc tacatttttt atgtttatct
     4621 ctagtattac tctttagaca aaaaaattgt agtaagaact attcatagag tgaatcgaaa
     4681 acaatacgaa aatgtaaaca tttcctatac gtagtatata gagacaaaat agaagaaacc
     4741 gttcataatt ttctgaccaa tgaagaatca tcaacgctat cactttctgt tcacaaagta
     4801 tgcgcaatcc acatcggtat agaatataat cggggatgcc tttatcttga aaaaatgcac
     4861 ccgcagcttc gctagtaatc agtaaacgcg ggaagtggag tcaggctttt tttatggaag
     4921 agaaaataga caccaaagta gccttcttct aaccttaacg gacctacagt gcaaaaagtt
     4981 atcaagagac tgcattatag agcgcacaaa ggagaaaaaa agtaatctaa gatgctttgt
     5041 tagaaaaata gcgctctcgg gatgcatttt tgtagaacaa aaaagaagta tagattcttt
     5101 gttggtaaaa tagcgctctc gcgttgcatt tctgttctgt aaaaatgcag ctcagattct
     5161 ttgtttgaaa aattagcgct ctcgcgttgc atttttgttt tacaaaaatg aagcacagat
     5221 tcttcgttgg taaaatagcg ctttcgcgtt gcatttctgt tctgtaaaaa tgcagctcag
     5281 attctttgtt tgaaaaatta gcgctctcgc gttgcatttt tgttctacaa aatgaagcac
     5341 agatgcttcg ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     6061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6181 ctggagccgg tgagcgtgga tctcgcggta tcattgcagc actggggcca gatggtaagc
     6241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7141 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7201 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7261 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7321 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7381 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7441 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7501 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     7561 tcagttta
//
