LOCUS       sequence-300756-        6768 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 6768)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6768)
  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-m4252883
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(11..57)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    75..123
                     /note="synthetic polyadenylation signal"
     enhancer        204..507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        509..711
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     662..682
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             915..935
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             936..1013
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1082..1203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1093..1112)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1147..1166
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1318..1364
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1394..1530
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1575..1646
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1872..2460)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1952..1971)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2960..4169
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4006..4025
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4173..5201
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5228..5276
                     /note="synthetic polyadenylation signal"
     CDS             join(5542..5610,5611..6402)
                     /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(5760..5779)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6415..6551
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(6703..6768,1..6)
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca
       61 atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt
      121 attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt
      181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      301 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      361 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      421 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      481 tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt
      541 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      601 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      661 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      721 aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg
      781 ccaccatgat agaactatcg ctcatcgact tttatctgtg cttcttggct ttcctgcttt
      841 ttctggtgct cattatgcta atcatttttt ggttcagtct ggagttacaa gatcataatg
      901 agacatgtca cgccgaaaac ctttactttc agtcgaagag gcgatggaaa aagaatttta
      961 tcgcggtgag tgcagccaac agattcaaaa agatatcctc gtctggtgcc ctctaaggat
     1021 ccgcaggcct ctgctagctt gactgactga gatacagcgt accttcagct cacagacatg
     1081 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1141 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1201 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1261 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     1321 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     1381 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     1441 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     1501 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     1561 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     1621 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     1681 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     1741 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     1801 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2041 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     2701 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     2761 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     2821 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     2881 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     2941 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     3001 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
     3061 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
     3121 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
     3181 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
     3241 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
     3301 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
     3361 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
     3421 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     3481 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     3541 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     3601 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     3661 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     3721 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     3781 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     3841 accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     3901 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     3961 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     4021 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4081 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     4141 tctggccgtt tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga
     4201 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     4261 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
     4321 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
     4381 ccgccgcgct cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct
     4441 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     4501 ccgctgttct gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc
     4561 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     4621 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     4681 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     4741 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     4801 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     4861 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     4921 tcgagcggag gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca
     4981 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     5041 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     5101 tcgcccgcag aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     5161 gaaaccgacg ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt
     5221 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     5281 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     5341 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     5401 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     5461 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     5521 ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt
     5581 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5641 gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5701 atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg
     5761 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5821 cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat
     5881 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5941 aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6001 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6061 gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact
     6121 ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa
     6181 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6241 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6301 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6361 cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg
     6421 gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt
     6481 cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa
     6541 tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     6601 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     6661 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     6721 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagcta
//
