LOCUS       sequence-307361-        7081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7081)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7081)
  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-m4252961
FEATURES             Location/Qualifiers
     source          1..7081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..608
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     559..579
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             813..833
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1182..1199
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1292..1413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1303..1322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1357..1376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1528..1574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1604..1740
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1785..1856
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2082..2670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2162..2181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3170..4379
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4216..4235
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4383..5411
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5438..5486
                     /note="synthetic polyadenylation signal"
     CDS             join(5752..5820,5821..6612)
                     /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(5970..5989)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6625..6761
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6913..6984
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6989..7035)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(7053..7081,1..20)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa
       61 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc
      421 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      481 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      541 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      601 gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac
      661 acacccgcca gcggcccgcc accatgatag aactatcgct catcgacttt tatctgtgct
      721 tcttggcttt cctgcttttt ctggtgctca ttatgctaat cattttttgg ttcagtctgg
      781 agttacaaga tcataatgag acatgtcacg ccgaaaacct ttactttcag tcgatgagtg
      841 acaaaatcat tcatctaacc gatgactcct ttgatacaga cgtgttgaag gccgatggtg
      901 cgatactggt agatttctgg gctgaatggt gtggcccgtg caagatgatc gcacccattc
      961 tggacgagat cgccgacgag tatcaaggaa aattaactgt cgctaagctt aatatagatc
     1021 agaacccagg gacggcacct aaatacggca tcaggggaat acctaccctc ctgctcttta
     1081 agaatgggga agttgccgcg acaaaagtgg gagccctgtc taagggccag ctaaaagagt
     1141 tcttggacgc taacctggca ggttcaggct cggggcacat gcaccatcac catcaccata
     1201 gcagcgggct cgtgccacgc ggctaaggat ccgcaggcct ctgctagctt gactgactga
     1261 gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt tggacaaacc
     1321 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1381 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1441 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     1501 ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct aaacgggtct
     1561 tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa
     1621 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     1681 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     1741 ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     1801 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     1861 cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga
     1921 tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg atctggtcct
     1981 tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc aaaaggccag
     2041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2881 tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag gccctcgtcg
     2941 gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa cgaactatcg
     3001 caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat cgccaggtat
     3061 tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac ctacatcctc
     3121 aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc
     3181 cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac
     3241 gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct
     3301 cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg
     3361 gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc
     3421 ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg
     3481 acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt
     3541 ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt
     3601 cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag
     3661 tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc
     3721 ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac
     3781 aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg
     3841 aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac gtgaagtttg
     3901 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     3961 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     4021 ttctgttggc ttataatgca gggtggggcc accagccggt aggtgtgcgg taggcttttc
     4081 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     4141 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     4201 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     4261 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     4321 agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagaca
     4381 acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4441 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4501 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4561 aagatcgtta tgtttatcgg cactttgcat ccgccgcgct cccgattccg gaagtgcttg
     4621 acattgggga gttcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4681 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcaa
     4741 tggatgccat cgctgccgcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4801 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4861 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4921 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4981 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5041 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5101 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5161 cgccccggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5221 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5281 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgccgcc gtctggaccg
     5341 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5401 caaaggaata agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     5461 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     5521 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     5581 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     5641 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     5701 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     5761 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5821 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     5881 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     5941 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     6001 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     6061 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     6121 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     6181 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     6241 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     6301 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6361 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6421 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6481 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6541 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6601 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     6661 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     6721 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     6781 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     6841 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     6901 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     6961 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     7021 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     7081 t
//
