LOCUS       sequence-307748-        4358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of 3a delta N(aa 42-275)-sfGFP, needs to be 
            introduced into baculoviral bacmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4358)
  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 4358)
  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_DK198
FEATURES             Location/Qualifiers
     source          1..4358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(125..713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(205..224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  903..1068
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    1189..1323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1213..1232)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1267..1286
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1456..1476)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             complement(1486..2199)
                     /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             complement(2245..2268)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     promoter        complement(3036..3127)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(3094..3111)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     protein_bind    3204..3237
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(3308..3313,3314..3330,3331..3336)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3525..4058
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4125..4348
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
ORIGIN
        1 cagggcatct gcgcatccac aggaagagcg acccaagtca atcaggggat aacgcaggaa
       61 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttgaccc aagtcaaccg
      901 gttgtgggcg gacaaaatag ttgggaactg ggaggggtgg aaatggagtt tttaaggatt
      961 atttagggaa gagtgacaaa atagatggga actgggtgta gcgtcgtaag ctaatacgaa
     1021 aattaaaaat gacaaaatag tttggaacta gatttcactt atctggttcc aattagatgg
     1081 gtatacccta ggggttatga tagttattgc tcagcggtgg cagcagccaa ctcagcttcc
     1141 tttcgggctt tgttagcagc cggatcttct aggctcaagc agtgatcaga tccagacatg
     1201 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1261 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1321 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1381 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatcc tctagtactt
     1441 ctcgacaagc tttcagtggt gatgatgatg atgatggctg ctgcctttgt acagttcatc
     1501 cataccatgc gtgatgcccg ctgcggttac gaactccagc agaaccatat gatcgcgttt
     1561 ctcgttcgga tctttagaca gaacgctttg cgtgctcaga tagtgattgt ctggcagcag
     1621 aacaggacca tcaccgattg gagtgttttg ctggtagtga tcagccagct gcacgctgcc
     1681 atcctccacg ttgtggcgaa ttttaaaatt cgctttaatg ccattttttt gtttatcggc
     1741 ggtgatgtaa acattgtggc tgttaaaatt gtattccagc ttatggccca ggatattgcc
     1801 gtcttcttta aagtcaatgc ctttcagctc aatgcggttt accagggtat cgccttcaaa
     1861 tttcacttcc gcacgcgttt tgtacgtgcc gtcatcctta aaggaaatcg tgcgttcctg
     1921 cacatagcct tccggcatgg cggacttgaa gaagtcatgc tgcttcatat ggtccggata
     1981 acgagcaaag cactgaacac cataagtcag cgtcgttacc agagtcggcc aaggtaccgg
     2041 cagtttacca gtagtacaga tgaacttcag cgtcagttta ccattagttg cgtcaccttc
     2101 accctcgcca cgcacggaaa acttatgacc gttgacatca ccatccagtt ccaccagaat
     2161 agggacgaca ccagtgaaca gctcttcgcc tttacgcata gaaccagaac cagatccagc
     2221 tgcagctcca gatccacctc tagacggacc ttgaaacaaa acttccaaag aattcgacag
     2281 ggggacggat gttgtcgtgg tgggttcgtc gtaaattggc tccataacgg ggttcacgac
     2341 accgctgcta ccgtcgatag tatgaatctg tacatgctcc tcgggttcgt ccacaatttt
     2401 attgtaaatg aaaaatgtta cgtgctcaac tcctgtatca gtagacaact gtgttgagta
     2461 gagttgataa taatccgagg taaaatatga gtgcagcact acacaatctt tgactccact
     2521 ctcccatttc tcggtgtagc cacctatttg gtaatcgtgt tcagaaatag gtgatgttgt
     2581 accgtcaccg gacgttatca cgattgaaga agtgacgcta ttatacggaa tacagtagtc
     2641 gtagcaattc gtatgccaac acaggaagta attggcatcg tacaaaagcg ggttcttgga
     2701 gcggcatttc caacaaagcc acagtctcat tattatcctg acaaaattta tgctctgcag
     2761 aaaatagacc agagcgtaca ggtaaaggaa gggagcttcc aggccagcag caacaagcaa
     2821 gagatgtgag taaacggtga caaagagcaa caggaggttg cacacgaagt gaacgccttt
     2881 tgagagcgcc agttgccaac gcttcttcag agtgattatc ttagaagctg actgaaagac
     2941 tgcaagcaac gcaacaccaa ctatcaacca accgaatggc atggtggctc gagggatccg
     3001 cgcccgatgg tgggacggta tgaataatcc ggaatattta taggtttttt tattacaaaa
     3061 ctgttacgaa aacagtaaaa tacttattta tttgcgagat ggttatcatt ttaattatct
     3121 ccatgatcta ttaatattcc ggagtaggtc gcgaatcgat actagtgttt aaactcgcta
     3181 ccttaggacc gttatagtta cagataactt cgtataatgt atgctatacg aagttatggc
     3241 gacttcgctg ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     3301 aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     3361 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     3421 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     3481 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca
     3541 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca
     3601 tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc
     3661 ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt
     3721 acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag
     3781 cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga
     3841 tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt gccaccgcgc
     3901 tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag
     3961 cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag
     4021 tgatgcactt tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg
     4081 gcttcccggc cgcggagttg ttcggtaaat tgtcacaacg ccgcgaatat agtctttaca
     4141 tgcccttggc cacgcccctc tttaatacga cgggcaattt gcacttcaga aaatgaagag
     4201 tttgctttag ccataacaaa agtccagtat gctttttcac agcataactg gactgatttc
     4261 agtttacaac tattctgtct agtttaagac tttattgtca tagtttagat ctattttgtt
     4321 cagtttaaga ctttattgtc cgcccacacc cgcttacg
//
