LOCUS       sequence-342381-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5052)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5052)
  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-m4133669
FEATURES             Location/Qualifiers
     source          1..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1408..1428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1429..1446
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1499..1620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1510..1529)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1564..1583
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1735..1781
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1811..1947
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1992..2063
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2289..2877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2369..2388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3623..3691,3692..4483)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGXRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3841..3860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4496..4632
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4784..4855
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4860..4906)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4924..4972
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggattt gtttatgaga
      601 atcttcacaa ttggaactgt aactttgaag caaggtgaaa tcaaggatgc tactccttca
      661 gattttgttc gcgctactgc aacgataccg atacaagcct cactcccttt cggatggctt
      721 attgttggcg ttgcacttct tgctgttttt cagagcgctt ccaaaatcat aaccctcaaa
      781 aagagatggc aactagcact ctccaagggt gttcactttg tttgcaactt gctgttgttg
      841 tttgtaacag tttactcaca ccttttgctc gttgctgctg gccttgaagc cccttttctc
      901 tatctttatg ctttagtcta cttcttgcag agtataaact ttgtaagaat aataatgagg
      961 ctttggcttt gctggaaatg ccgttccaaa aacccattac tttatgatgc caactatttt
     1021 ctttgctggc atactaattg ttacgactat tgtatacctt acaatagtgt aacttcttca
     1081 attgtcatta cttcaggtga tggcacaaca agtcctattt ctgaacatga ctaccagatt
     1141 ggtggttata ctgaaaaatg ggaatctgga gtaaaagact gtgttgtatt acacagttac
     1201 ttcacttcag actattacca gctgtactca actcaattga gtacagacac tggtgttgaa
     1261 catgttacct tcttcatcta caataaaatt gttgatgagc ctgaagaaca tgtccaaatt
     1321 cacacaatcg acggttcatc cggagttgtt aatccagtaa tggaaccaat ttatgatgaa
     1381 ccgacgacga ctactagcgt gcctttggaa aacctttact ttcagtcgca ccatcaccat
     1441 caccattaag ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
     1501 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1561 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1621 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1681 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
     1741 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
     1801 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
     1861 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
     1921 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
     1981 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
     2041 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
     2101 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
     2161 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
     2221 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     2281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     3121 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     3181 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     3241 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     3301 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     3361 aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     3421 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     3481 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     3541 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     3601 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3661 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3721 tgctgaagat cagttgggtg crcgagtggg ttacatcgaa ctggatctca acagcggtaa
     3781 gatccttgag agttttcgcc ccgaagaacg ytttccaatg atgagcactt ttaaagttct
     3841 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3901 acactattct cagaatgact tggttgagta ytcaccagtc acagaaaagc atcttacgga
     3961 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4021 caacttactt ctgacaacga tyggaggacc gaaggagcta accgcttttt tgcacaacat
     4081 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4141 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acsttgcgya aactattaac
     4201 tggcgaacta cttactctag cttcccggca acarttrata gactggatgg aggcggataa
     4261 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4321 tggagccggt gagcgtggrw sycgcggtat cattgcagca ctggggccag atggtaagcc
     4381 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4441 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     4501 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     4561 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     4621 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     4681 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     4741 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     4801 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     4861 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     4921 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     4981 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     5041 tatggagttc cg
//
