LOCUS       sequence-307329-        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-m4252880
FEATURES             Location/Qualifiers
     source          1..6768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             132..152
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             153..230
                     /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    299..420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(310..329)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     364..383
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      535..581
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      611..747
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    792..863
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1089..1677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1169..1188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2177..3386
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3223..3242
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3390..4418
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4445..4493
                     /note="synthetic polyadenylation signal"
     CDS             join(4759..4827,4828..5619)
                     /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(4977..4996)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5632..5768
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5920..5991
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5996..6042)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6060..6108
                     /note="synthetic polyadenylation signal"
     enhancer        6189..6492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6494..6696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6647..6667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ccatgattga actttcatta attgacttct atttgtgctt tttagccttt ctgctattcc
       61 ttgttttaat tatgcttatt atcttttggt tctcacttga actgcaagat cataatgaaa
      121 cttgtcacgc cgaaaacctt tactttcagt cgaagaggcg atggaaaaag aattttatcg
      181 cggtgagtgc agccaacaga ttcaaaaaga tatcctcgtc tggtgccctc taaggatccg
      241 caggcctctg ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
      301 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
      361 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
      421 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
      481 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
      541 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
      601 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
      661 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
      721 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
      781 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
      841 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
      901 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
      961 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     1021 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1501 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1741 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1801 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1861 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     1921 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     1981 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     2041 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     2101 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     2161 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     2221 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     2281 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     2341 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     2401 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     2461 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     2521 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     2581 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     2641 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     2701 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     2761 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     2821 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     2881 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     2941 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     3001 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     3061 agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     3121 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     3181 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     3241 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     3301 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     3361 ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt
     3421 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg
     3481 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
     3541 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg
     3601 ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
     3661 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     3721 ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga
     3781 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     3841 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     3901 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     3961 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     4021 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     4081 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     4141 agcggaggca tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg
     4201 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     4261 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     4321 cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
     4381 accgacgccc cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt
     4441 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     4501 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     4561 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     4621 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     4681 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     4741 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4801 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4861 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4921 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     4981 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5041 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     5101 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5161 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5221 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5281 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     5341 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     5401 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5461 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5521 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5581 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     5641 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     5701 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     5761 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     5821 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     5881 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     5941 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     6001 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     6061 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     6121 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     6181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6481 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6661 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     6721 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgcca
//
