LOCUS       62056_13770        7564 bp DNA     circular SYN 29-MAY-2021
DEFINITION  Cloning vector pLDJIF20, complete sequence.
ACCESSION   MW820852
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7564)
  AUTHORS   Ding L, Brown DM, Glass JI.
  TITLE     Rescue of Infectious Sindbis Virus by Yeast Spheroplast-Mammalian 
            Cell Fusion
  JOURNAL   Viruses 13 (4), 603 (2021)
  PUBMED    33916100
REFERENCE   2  (bases 1 to 7564)
  AUTHORS   Ding L.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   3  (bases 1 to 7564)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Viruses"; 
            date: "2021"; volume: "13"; issue: "4"; pages: "603"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-MAR-2021) Synthetic Biology "
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     misc_feature    443..472
                     /note="Harvard overlap; adapter 1"
     misc_feature    479..484
                     /label=yeast translational entry site
                     /note="yeast translational entry site"
     CDS             485..514
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     misc_feature    515..517
                     /label=yeast translational stop codon
                     /note="yeast translational stop codon"
     misc_feature    518..547
                     /note="Harvard overlap; adapter 2"
     terminator      548..735
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     promoter        760..1403
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate
                     dehydrogenase; also known as the TDH3 promoter"
     CDS             1421..2128
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     CDS             2156..2863
                     /codon_start=1
                     /transl_table=11
                     /product="yeCherry"
                     /label=yeCherry
                     /note="red fluorescence protein; 2nd copy"
                     /protein_id="QVS03008.1"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     terminator      2870..3117
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3418..4006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4180..4992)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        complement(4993..5097)
                     /label=AmpR promoter
     misc_feature    5134..5637
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     promoter        5893..6173
                     /label=TRP1 promoter
     CDS             6174..6974
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      complement(7074..7529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7564
                     /label=GAL1
                     /note="Saccharomyces cerevisiae galactokinase promoter"
ORIGIN
        1 cggattagaa gccgccgagc gggtgacagc cctccgaagg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa tgaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatgcaaa aactgcataa ccactttaac taatactttc aacattttcg
      361 gtttgtatta cttcttattc aaatgtaata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agagaaccca ctgcttactg gcttatcgaa ataaaaaaat
      481 gtctgaacaa aaactcatct cagaagagga tctgtaactg tgccttctag ttgccagcca
      541 tctgttggcg aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat
      601 aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag
      661 taactctttc ctgtaggtca ggttgctttc tcaggtatag tatgaggtcg ctcttattga
      721 ccacacctct accggaacgg gtgcgcatag aaatgtttat cattatcaat actcgccatt
      781 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
      841 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
      901 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
      961 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     1021 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     1081 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     1141 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     1201 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     1261 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     1321 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     1381 aaaacaccag aacttagttt cgacggattc tagaactagt atggtttcaa agggcgaaga
     1441 agacaatatg gctattatta aggaattcat gagattcaag gtccacatgg aaggttctgt
     1501 caacggtcac gaattcgaaa ttgaaggtga aggtgaaggt agaccatacg aaggtaccca
     1561 aaccgctaag ttgaaggtca ccaagggtgg tccattgcca ttcgcttggg atattttgtc
     1621 tccacaattc atgtacggtt ctaaggctta cgtcaagcac ccagctgata ttccagatta
     1681 cttgaagttg tctttcccag aaggtttcaa gtgggaaaga gtcatgaact tcgaagatgg
     1741 tggtgtcgtc accgtcaccc aagattcttc tttgcaagat ggtgaattca tttacaaggt
     1801 caagttgaga ggtaccaact tcccatctga tggtccagtc atgcaaaaga agaccatggg
     1861 ttgggaagct tcttctgaaa gaatgtaccc agaagatggt gctttgaagg gtgaaattaa
     1921 gcaaagattg aagttgaagg atggtggtca ctacgatgct gaagtcaaga ccacctacaa
     1981 ggctaagaag ccagtccaat tgccaggtgc ttacaacgtc aacattaagt tggatattac
     2041 ctctcacaac gaagattaca ccattgtcga acaatacgaa agagctgaag gtagacactc
     2101 taccggtggt atggatgaat tgtacaagta cctcgagtgc ggccgctacg ctagcgtttc
     2161 aaagggcgaa gaagacaata tggctattat taaggaattc atgagattca aggtccacat
     2221 ggaaggttct gtcaacggtc acgaattcga aattgaaggt gaaggtgaag gtagaccata
     2281 cgaaggtacc caaaccgcta agttgaaggt caccaagggt ggtccattgc cattcgcttg
     2341 ggatattttg tctccacaat tcatgtacgg ttctaaggct tacgtcaagc acccagctga
     2401 tattccagat tacttgaagt tgtctttccc agaaggtttc aagtgggaaa gagtcatgaa
     2461 cttcgaagat ggtggtgtcg tcaccgtcac ccaagattct tctttgcaag atggtgaatt
     2521 catttacaag gtcaagttga gaggtaccaa cttcccatct gatggtccag tcatgcaaaa
     2581 gaagaccatg ggttgggaag cttcttctga aagaatgtac ccagaagatg gtgctttgaa
     2641 gggtgaaatt aagcaaagat tgaagttgaa ggatggtggt cactacgatg ctgaagtcaa
     2701 gaccacctac aaggctaaga agccagtcca attgccaggt gcttacaacg tcaacattaa
     2761 gttggatatt acctctcaca acgaagatta caccattgtc gaacaatacg aaagagctga
     2821 aggtagacac tctaccggtg gtatggatga attgtacaag taactcgagt catgtaatta
     2881 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     2941 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     3001 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     3061 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcccc
     3121 gggcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3181 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3241 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3421 ccataggctc ggcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3481 aaacccgaca ggactataaa gataccaggc gttcccccct ggaagctccc tcgtgcgctc
     3541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3601 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3841 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4081 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4141 aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa
     4201 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     4261 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     4321 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     4381 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     4441 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     4501 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     4561 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     4621 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     4681 ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     4741 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     4801 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     4861 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     4921 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     4981 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5101 atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat
     5161 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     5221 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     5281 accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     5341 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     5401 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     5461 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     5521 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     5581 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt
     5641 cctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5701 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5761 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5821 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5881 gagtgcacca taaacgacat tactatatat ataatatagg aagcatttaa tagacagcat
     5941 cgtaatatat gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt
     6001 attgaaaaat agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc
     6061 gattaagaat taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt
     6121 gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtcga
     6181 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     6241 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     6301 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     6361 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     6421 agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat
     6481 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     6541 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     6601 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     6661 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     6721 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     6781 gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg
     6841 tggtctctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     6901 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     6961 gcggccagca aaactaagtt attactgagt agtatttatt taagtattgt ttgtgcactt
     7021 gcctgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     7081 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7141 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7201 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7261 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7321 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7381 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7441 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7501 cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg ccattcaggc tgcgcaactg
     7561 ttgg
//
