LOCUS       sequence-307332-        4849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4849)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4849)
  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-m4046651
FEATURES             Location/Qualifiers
     source          1..4849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        64..477
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(664..682)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1033..1836
                     /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      1919..2901
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2979..3008
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3030..3617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3629..3656)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3680..3871)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3872..4663,4664..4732))
                     /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     4495..4514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4733..4835)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(4836..4849,1..18))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
       61 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
      121 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      181 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      241 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      301 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      361 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      421 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      481 taattacaaa atgatcgagt tatccttgat agatttctat ctatgttttt tggcattcct
      541 gctttttctc gtgttgatta tgttgattat cttctggttc agtttagaat tgcaagacca
      601 taatgaaaca tgccacgcct aaacaggccc cttttccttt gtcgatatca tgtaattagt
      661 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
      721 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
      781 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
      841 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
      901 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
      961 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     1021 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     1081 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     1141 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     1201 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     1261 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     1321 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     1381 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     1441 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     1501 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     1561 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     1621 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     1681 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     1741 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     1801 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     1861 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     1921 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     1981 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     2041 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     2101 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     2161 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     2221 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     2281 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     2341 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     2401 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     2461 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     2521 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     2581 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     2641 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     2701 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     2761 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     2821 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     2881 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     2941 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     3001 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3121 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3181 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3241 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3301 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3361 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3421 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3481 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     3541 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     3601 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     3661 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     3721 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     3781 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     3841 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     3901 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3961 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4021 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     4081 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4141 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4201 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4261 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4321 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4381 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     4441 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4501 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     4561 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4621 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4681 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     4741 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     4801 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctcc
//
