LOCUS       sequence-307323-        5977 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast expression plasmid for SARS-CoV-2 N (nucleocapsid).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5977)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046696
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(203..214)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1315..1333)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1684..2487
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2570..3552
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3630..3659
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3681..4268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4280..4307)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4331..4522)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4523..5314,5315..5383))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5146..5165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5384..5486)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5487..5518)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        5564..5977
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ttttaattac aaaatgtctg ataatggccc acagaaccaa cgtaacgcac ctagaatcac
       61 cttcggtggt ccatccgaca gcactggttc aaatcaaaac ggagagagat cgggtgcccg
      121 gagtaagcaa aggcgccccc aaggtctacc gaataacaca gcttcttggt ttactgctct
      181 tacgcaacac ggtaaagaag acttgaagtt cccaagaggt caaggggttc caattaacac
      241 caactcctct ccagatgatc aaataggtta ctatagaaga gctactagac gtattagggg
      301 tggggacggc aaaatgaagg atttatctcc tagatggtac ttttattact tgggtaccgg
      361 tccagaagct ggattgcctt acggtgccaa taaggacggt atcatttggg tagcaactga
      421 aggtgcttta aataccccaa aggaccatat cggtactaga aaccctgcta ataacgctgc
      481 cattgtcttg caattaccac aggggacaac gttgccaaag ggtttctacg ccgaaggatc
      541 cagaggtggc tcgcaagctt ctagtagatc atcctctcgc agcagaaact catctcgtaa
      601 ctccacacca ggtagttcga gaggtacctc tccagcgaga atggctggta acggtggtga
      661 tgcagctctg gcgttgctgc tactcgacag attgaatcaa ttggaatcca aaatgtctgg
      721 taagggtcaa caacaacagg gtcagactgt gactaagaag tctgctgctg aagcttccaa
      781 aaagccccgt caaaagagaa ccgccaccaa ggcctataat gtcactcaag ctttcggaag
      841 aagaggtcca gagcaaacgc aaggtaactt tggagatcaa gaattgatcc ggcagggtac
      901 cgattataaa cactggcccc aaattgctca attcgcgcca agcgcctcag cttttttcgg
      961 catgtcaaga ataggtatgg aagttactcc atctgggact tggttaactt acactggtgc
     1021 aatcaagttg gacgataaag acccaaattt taaagaccaa gttattctac tcaacaagca
     1081 tatagatgct tacaaaacct tcccgcctac agaacccaag aaagataaaa agaagaaggc
     1141 tgacgagaca caagcattgc ctcagcgtca aaagaaacaa caaactgtca ccttgttacc
     1201 agcagctgat cttgatgatt tctctaagca attgcaacaa tccatgtcgt ccgccgactc
     1261 tactcaagcg taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     1321 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     1381 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     1441 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     1501 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     1561 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     1621 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     1681 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     1741 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     1801 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     1861 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     1921 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     1981 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     2041 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     2101 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     2161 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     2221 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     2281 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     2341 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     2401 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     2461 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     2521 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     2581 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     2641 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     2701 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     2761 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     2821 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     2881 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     2941 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     3001 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     3061 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     3121 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     3181 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     3241 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     3301 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     3361 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     3421 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     3481 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     3541 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     3601 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     3661 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3781 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4081 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4141 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     4201 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4261 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     4321 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4381 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4441 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4501 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     4561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     4621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4681 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     4741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     4981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5041 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     5221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5341 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     5401 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     5461 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     5521 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga gcggccgccc cacacaccat
     5581 agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc
     5641 gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta
     5701 gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc
     5761 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt
     5821 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
     5881 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
     5941 tgctcattag aaagaaagca tagcaatcta atctaag
//
