LOCUS       sequence-307746-        4478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of 3a(Q57H)-sfGFP, needs to be introduced 
            into baculoviral bacmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4478)
  AUTHORS   Kern DM, Sorum B, Mali SS, Hoel CM, Sridharan S, Remis JP, Toso DB, 
            Kotecha A, Bautista DM, Brohawn SG
  TITLE     Cryo-EM structure of the SARS-CoV-2 3a ion channel in lipid 
            nanodiscs.
  JOURNAL   bioRxiv [Preprint]. 2021 Jan 26:2020.06.17.156554. doi: 
            10.1101/2020.06.17.156554.
  PUBMED    32587976
REFERENCE   2  (bases 1 to 4478)
  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"; journalName: "bioRxiv 
            [Preprint]. 2021 Jan 26:2020.06.17.156554. doi: 
            10.1101/2020.06.17.156554."
COMMENT     Sequence Label: pSBL_DK193
FEATURES             Location/Qualifiers
     source          1..4478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     395..414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  629..852
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(919..1452)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1641..1646,1647..1663,1664..1669))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1740..1773)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1850..1941
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1866..1883
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      1989..1998
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2829..2852
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2898..3611
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3621..3641
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3774..3908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3811..3830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3865..3884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4029..4194
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(4384..4478,1..494)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
       61 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      541 cttttgctca catgttcttt cctgcgttat cccctgattg acttgggtcg ctcttcctgt
      601 ggatgcgcag atgccctgcg taagcgggtg tgggcggaca ataaagtctt aaactgaaca
      661 aaatagatct aaactatgac aataaagtct taaactagac agaatagttg taaactgaaa
      721 tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa agcaaactct
      781 tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca agggcatgta
      841 aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat
      901 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
      961 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
     1021 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
     1081 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
     1141 actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
     1201 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
     1261 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
     1321 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
     1381 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     1441 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     1501 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     1561 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     1621 gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg
     1681 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgcca
     1741 taacttcgta tagcatacat tatacgaagt tatctgtaac tataacggtc ctaaggtagc
     1801 gagtttaaac actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
     1861 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     1921 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     1981 tccctcgagc caccatggat ctctttatgc gcatttttac gatcggtaca gttacgttga
     2041 agcaaggtga aatcaaagac gccacgccgt ccgattttgt tcgcgctacg gctacaattc
     2101 caatccaagc gagcttgcca ttcggttggt tgatagttgg tgttgcgttg cttgcagtct
     2161 ttcactcagc ttctaagata atcactctga agaagcgttg gcaactggcg ctctcaaaag
     2221 gcgttcactt cgtgtgcaac ctcctgttgc tctttgtcac cgtttactca catctcttgc
     2281 ttgttgctgc tggcctggaa gctcccttcc tttacctgta cgctctggtc tattttctgc
     2341 agagcataaa ttttgtcagg ataataatga gactgtggct ttgttggaaa tgccgctcca
     2401 agaacccgct tttgtacgat gccaattact tcctgtgttg gcatacgaat tgctacgact
     2461 actgtattcc gtataatagc gtcacttctt caatcgtgat aacgtccggt gacggtacaa
     2521 catcacctat ttctgaacac gattaccaaa taggtggcta caccgagaaa tgggagagtg
     2581 gagtcaaaga ttgtgtagtg ctgcactcat attttacctc ggattattat caactctact
     2641 caacacagtt gtctactgat acaggagttg agcacgtaac atttttcatt tacaataaaa
     2701 ttgtggacga acccgaggag catgtacaga ttcatactat cgacggtagc agcggtgtcg
     2761 tgaaccccgt tatggagcca atttacgacg aacccaccac gacaacatcc gtccccctgt
     2821 cgaattcttt ggaagttttg tttcaaggtc cgtctagagg tggatctgga gctgcagctg
     2881 gatctggttc tggttctatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc
     2941 tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag
     3001 gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg
     3061 taccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc
     3121 cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg
     3181 aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg
     3241 aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca
     3301 atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg
     3361 ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca
     3421 gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc
     3481 tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac
     3541 gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg
     3601 aactgtacaa aggcagcagc catcatcatc atcatcacca ctgaaagctt gtcgagaagt
     3661 actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3721 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3781 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3841 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3901 tctggatctg atcactgctt gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa
     3961 gctgagttgg ctgctgccac cgctgagcaa taactatcat aacccctagg gtatacccat
     4021 ctaattggaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     4081 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     4141 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacaaccggt
     4201 tgacttgggt caactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4261 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4321 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4381 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4441 accagcggtg gtttgtttgc cggatcaaga gctaccaa
//
