LOCUS       sequence-306273-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for Human coronavirus HKU1 
            nucleocapsid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  TITLE     Human coronavirus mammalian expression constructs
REFERENCE   2  (bases 1 to 7418)
  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-m4134897
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1380..1501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1391..1410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1445..1464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1616..1662
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1692..1828
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1873..1944
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2170..2758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2250..2269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3258..4467
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4304..4323
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4471..5070
                     /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(4471..4490)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4967..4987
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5097..5145
                     /note="synthetic polyadenylation signal"
     CDS             join(5411..5479,5480..6271)
                     /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(5629..5648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6284..6420
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6572..6643
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6648..6694)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6712..6760
                     /note="synthetic polyadenylation signal"
     enhancer        6841..7144
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7146..7348
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7299..7319
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtct tatactcccg gtcattatgc tggaagtaga agctcctctg gaaatcgttc
       61 aggaatcctc aagaaaactt cttgggctga ccaatctgag cgaaattacc aaacctttaa
      121 tagaggcaga aaaacccaac ctaaattcac tgtgtctact caaccacaag gaaatactat
      181 cccacattat tcctggttct ccgggatcac tcaatttcaa aaaggtagag actttaaatt
      241 ttcagatggt caaggagttc ccattgcttt cggagtaccc ccttctgaag caaaaggata
      301 ttggtataga cacagccggc gttcttttaa aacagctgat ggtcaacaaa agcagttgtt
      361 accgagatgg tatttctact atctcggtac cggcccatat gccaatgcat cctatggtga
      421 atccctcgaa ggggtcttct gggttgctaa tcaccaagct gacacttcta ctccctccga
      481 tgtttcgtca agggatccta ctactcaaga agctatccct actaggtttc cgcctggtac
      541 gattttgcct caaggctatt atgttgaagg ctcaggaagg tctgcttcta atagtcgacc
      601 aggttcacgt tctcaatcac gtggacccaa taatcgttca ttaagtagaa gtaattctaa
      661 ttttagacat tcagattcta tagtaaaacc tgatatggct gatgagatcg ctaatcttgt
      721 tttagccaag cttggtaaag attctaaacc tcagcaagtc actaagcaaa atgccaagga
      781 aatcaggcat aaaattttaa caaaacctcg ccaaaagcga actcctaata aacattgtaa
      841 tgttcaacag tgttttggta aaagaggacc ttctcaaaat tttggtaatg ctgaaatgtt
      901 aaagcttggt actaatgatc ctcagtttcc tattcttgca gaattagctc ctacaccagg
      961 tgcttttttc tttggttcta aattagactt ggttaaaaga gattccgagg ctgactcacc
     1021 tgttaaagat gtttttgaac ttcattattc tggttctatt aggtttgata gtactttacc
     1081 aggctttgag acaattatga aagttcttga agagaattta aatgcttacg ttaattctaa
     1141 tcagaacact gattctgatt cgttgagttc taaacctcag cgtaaaagag gtgttaaaca
     1201 attaccagaa cagtttgact ctcttaattt aagtgctggt actcagcaca tttcaaatga
     1261 ttttactcct gaggatcata gtttacttgc tactcttgat gatccttatg tagaagactc
     1321 tgttgcttaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1381 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1441 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1501 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1561 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1621 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1681 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1741 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1801 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1861 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1921 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1981 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     2041 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     2101 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2341 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2581 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     3001 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     3061 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     3121 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     3181 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3241 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3301 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     3361 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     3421 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     3481 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     3541 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     3601 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     3661 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3721 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     3781 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3841 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3901 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3961 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     4021 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     4081 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     4141 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     4201 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     4261 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4321 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4381 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     4441 tggccgtttt tggctttttt gttagacaac atgaccgagt acaagcccac ggtgcgcctc
     4501 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4561 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4621 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4681 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4741 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggag
     4801 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4861 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcc
     4921 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4981 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5041 tggtgcatga cccgcaagcc cggtgcctag gctagtatgt aagcctagtc ttagataata
     5101 aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa cgctaggcgc
     5161 gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata tcgttgcttg
     5221 gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca gatattgtac
     5281 tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat agtctgatcg
     5341 gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag gatcagcagg
     5401 aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5521 gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5581 ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5701 gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg gcatgacagt
     5761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5821 gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5941 caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg gcgaactact
     6001 tactctagct tcccggcaac agttgataga ctggatggag gcggataaag ttgcaggacc
     6061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6241 gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg cgcagaaaaa
     6301 aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc agcaacggat
     6361 acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat ttatccttaa
     6421 gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     6481 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     6541 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     6601 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     6661 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     6721 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     6781 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6841 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6901 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     6961 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7021 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     7081 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     7141 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     7201 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     7261 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     7321 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     7381 gatcctacca tccactcgac acacccgcca gcggccgc
//
