LOCUS       sequence-300220-        7439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for Human coronavirus OC43 
            nucleocapsid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  TITLE     Human coronavirus mammalian expression constructs
REFERENCE   2  (bases 1 to 7439)
  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-m4134899
FEATURES             Location/Qualifiers
     source          1..7439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1401..1522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1412..1431)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1466..1485
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1637..1683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1713..1849
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1894..1965
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2191..2779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2271..2290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3279..4488
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4325..4344
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4492..5091
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4492..4511)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4988..5008
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5118..5166
                     /note="synthetic polyadenylation signal"
     CDS             join(5432..5500,5501..6292)
                     /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(5650..5669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6305..6441
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6593..6664
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6669..6715)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6733..6781
                     /note="synthetic polyadenylation signal"
     enhancer        6862..7165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7167..7369
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7320..7340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct tttactcctg gtaagcaatc cagtagtaga gcgtcctctg gaaatcgttc
       61 tggtaatggc atcctcaagt gggccgatca gtccgaccag gtcagaaatg ttcaaaccag
      121 gggtagaaga gctcaaccca agcaaactgc tacctctcag caaccatcag gagggaatgt
      181 tgtaccctac tattcttggt tctctggaat tactcagttt caaaagggaa aggagtttga
      241 gtttgtagaa ggacaaggtc cgcctattgc accaggagtc ccagctactg aagctaaggg
      301 gtactggtac agacacaaca gaggttcttt taaaacagcc gatggcaacc agcgtcaact
      361 gctgccacga tggtattttt actatctggg aacaggaccg catgctaaag accagtacgg
      421 caccgatatt gacggagtct actgggtcgc tagcaaccag gctgatgtca ataccccggc
      481 tgacattgtc gatcgggacc caagtagcga tgaggctatt ccgactaggt ttccgcctgg
      541 cacggtactc cctcagggtt actatattga aggctcagga aggtctgctc ctaattccag
      601 atctacttcg cgcacatcca gcagagcctc tagtgcagga tcgcgtagta gagccaattc
      661 tggcaataga acccctacct ctggtgtaac acctgacatg gctgatcaaa ttgctagtct
      721 tgttctggca aaacttggca aggatgccac taaacctcag caagtaacta agcatactgc
      781 caaagaagtc agacagaaaa ttttgaataa gccccgccag aagaggagcc ccaataaaca
      841 atgcactgtt cagcagtgtt ttggtaagag aggccctaat cagaattttg gtggtggaga
      901 aatgttaaaa cttggaacta gtgacccaca gttccccatt cttgcagaac tcgcacccac
      961 agctggtgcg tttttctttg gatcaagatt agagttggcc aaagtgcaga atttatctgg
     1021 gaatcctgac gagccccaga aggatgttta tgaattgcgc tataacggcg caattaggtt
     1081 tgacagtaca ctttcaggtt ttgagaccat aatgaaggtg ctgaatgaga atttgaatgc
     1141 ctatcaacaa caagatggta tgatgaatat gagtccaaaa ccacagcgtc agcgtggtca
     1201 taagaatgga caaggagaaa atgataatat aagtgttgca gtgcccaaaa gccgcgtgca
     1261 gcaaaataag agtagagagt tgactgcaga ggacatcagc cttcttaaga agatggatga
     1321 gccctatact gaagacacct cagaaatata agctagcttg actgactgag atacagcgta
     1381 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     1441 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     1501 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     1561 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
     1621 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1681 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
     1741 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     1801 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct
     1861 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
     1921 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
     1981 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
     2041 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     2101 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     2161 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2221 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2281 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2341 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3001 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     3061 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     3121 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     3181 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     3241 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     3301 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     3361 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     3421 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     3481 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     3541 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     3601 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     3661 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     3721 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     3781 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     3841 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     3901 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     3961 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     4021 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     4081 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     4141 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     4201 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     4261 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     4321 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     4381 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     4441 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     4501 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4561 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     4621 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4681 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4741 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4801 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4861 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4921 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4981 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5041 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     5101 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     5161 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5221 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5281 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5341 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     5401 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     5461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5521 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     5581 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     5641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5701 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     5761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5821 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     5881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     6001 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     6301 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     6361 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     6421 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     6481 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     6541 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     6601 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     6661 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     6721 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     6781 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     6841 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     6901 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6961 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7021 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     7081 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     7141 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     7201 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     7261 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     7321 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     7381 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
