LOCUS       sequence-287645-        5950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5950)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5950)
  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"
COMMENT     Sequence Label: pGBW-m4046490
FEATURES             Location/Qualifiers
     source          1..5950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             545..562
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1705..1723)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2074..2877
                     /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      2960..3942
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4020..4049
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4071..4658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4670..4697)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4721..4912)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4913..5704,5705..5773))
                     /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     5536..5555
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5774..5876)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5877..5908)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgggttaca taaatgtctt tgcattccct ttcaccatct atagtcttct
      481 attatgcaga atgaactccc gaaactacat tgcccaggtt gacgtagtta actttaattt
      541 gactctcgtg ccgagaggtt ctaaaatcga ggaaggtaag ctagtcattt ggataaatgg
      601 agataagggt tacaacggtt tggcagaagt aggcaagaaa tttgaaaagg acacaggtat
      661 caaggtgacg gttgaacatc cagacaagtt agaagaaaaa ttccctcaag ttgctgctac
      721 tggtgatggt cccgacatta ttttctgggc ccacgatcgt tttggtggat atgctcagtc
      781 aggtttgctg gctgagatca ccccagacaa ggccttccaa gataaattgt acccattcac
      841 ttgggatgct gtcagataca acggtaagct tatagcatat ccaattgctg ttgaagcctt
      901 atcgttgatc tacaataagg acttgttacc taacccacca aagacatggg aagaaattcc
      961 agctttggat aaagagctaa aggccaaggg aaaaagcgct ttgatgttta acttacaaga
     1021 accttatttc acctggccat taatcgctgc tgacggtggt tacgcattta aatacgaaaa
     1081 cggtaagtat gacattaagg atgtcggcgt tgataatgct ggtgcaaagg ctggtttgac
     1141 cttcttggtt gatttaataa agaataaaca catgaacgct gacactgact actctatcgc
     1201 cgaagccgct ttcaacaagg gtgaaactgc tatgaccata aatggtccat gggcctggag
     1261 taacattgat acttctaaag taaactatgg tgtcactgtt ttgcctacat tcaaaggcca
     1321 accatccaag ccctttgtcg gtgttctatc cgctggcatc aatgctgctt ctccaaacaa
     1381 ggagctagct aaagaatttt tggaaaatta cttgttaact gacgaaggtt tggaggccgt
     1441 taacaaggac aagccattgg gtgctgtcgc tttgaaatct tacgaagaag aattggctaa
     1501 ggatcccaga attgccgcta ccatggagaa cgcccaaaag ggtgaaatta tgccaaacat
     1561 cccacaaatg tcagccttct ggtacgcagt taggactgcc gtgattaatg ctgcttccgg
     1621 tagacaaacc gtcgacgaag cattaaaaga tgctcaaacg taaacaggcc ccttttcctt
     1681 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
     1741 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
     1801 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     1861 aaagcggccg ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
     1921 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     1981 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     2041 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     2101 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     2161 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     2221 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     2281 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     2341 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     2401 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     2461 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     2521 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     2581 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     2641 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     2701 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     2761 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     2821 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     2881 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     2941 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     3001 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     3061 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     3121 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     3181 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     3241 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     3301 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     3361 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     3421 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     3481 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     3541 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     3601 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     3661 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     3721 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     3781 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     3841 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     3901 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     3961 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     4021 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     4081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4141 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     4201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4561 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     4621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     4681 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     4741 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4801 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4861 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     4921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4981 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     5041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5101 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     5161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5341 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     5401 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     5461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5581 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5761 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     5821 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     5881 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     5941 gtcagttaga
//
