LOCUS       sequence-307747-        4478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of 3a-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_DK187
FEATURES             Location/Qualifiers
     source          1..4478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     302..321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  536..759
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(826..1359)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1548..1553,1554..1570,1571..1576))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1647..1680)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1757..1848
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1773..1790
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      1896..1905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2736..2759
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2805..3518
                     /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             3528..3548
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3681..3815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3718..3737)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3772..3791
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3936..4101
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(4291..4478,1..401)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
       61 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      121 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      181 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      241 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      301 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      361 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      421 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      481 ctgattgact tgggtcgctc ttcctgtgga tgcgcagatg ccctgcgtaa gcgggtgtgg
      541 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
      601 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
      661 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
      721 aaagaggggc gtggccaagg gcatgtaaag actatattcg cggcgttgtg acaatttacc
      781 gaacaactcc gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact
      841 tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac
      901 tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc
      961 ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga
     1021 gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta
     1081 agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt
     1141 actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac
     1201 gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg
     1261 gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg
     1321 actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca
     1381 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa
     1441 aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg
     1501 tcaaggttct ggaccagttg cgtgagcgca tacgctactt gcattacagt ttacgaaccg
     1561 aacaggctta tgtcaactgg gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg
     1621 caaccttggg cagcagcgaa gtcgccataa cttcgtatag catacattat acgaagttat
     1681 ctgtaactat aacggtccta aggtagcgag tttaaacact agtatcgatt cgcgacctac
     1741 tccggaatat taatagatca tggagataat taaaatgata accatctcgc aaataaataa
     1801 gtattttact gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat
     1861 tcataccgtc ccaccatcgg gcgcggatcc ctcgagccac catggatctc tttatgcgca
     1921 tttttacgat cggtacagtt acgttgaagc aaggtgaaat caaagacgcc acgccgtccg
     1981 attttgttcg cgctacggct acaattccaa tccaagcgag cttgccattc ggttggttga
     2041 tagttggtgt tgcgttgctt gcagtctttc agtcagcttc taagataatc actctgaaga
     2101 agcgttggca actggcgctc tcaaaaggcg ttcacttcgt gtgcaacctc ctgttgctct
     2161 ttgtcaccgt ttactcacat ctcttgcttg ttgctgctgg cctggaagct cccttccttt
     2221 acctgtacgc tctggtctat tttctgcaga gcataaattt tgtcaggata ataatgagac
     2281 tgtggctttg ttggaaatgc cgctccaaga acccgctttt gtacgatgcc aattacttcc
     2341 tgtgttggca tacgaattgc tacgactact gtattccgta taatagcgtc acttcttcaa
     2401 tcgtgataac gtccggtgac ggtacaacat cacctatttc tgaacacgat taccaaatag
     2461 gtggctacac cgagaaatgg gagagtggag tcaaagattg tgtagtgctg cactcatatt
     2521 ttacctcgga ttattatcaa ctctactcaa cacagttgtc tactgataca ggagttgagc
     2581 acgtaacatt tttcatttac aataaaattg tggacgaacc cgaggagcat gtacagattc
     2641 atactatcga cggtagcagc ggtgtcgtga accccgttat ggagccaatt tacgacgaac
     2701 ccaccacgac aacatccgtc cccctgtcga attctttgga agttttgttt caaggtccgt
     2761 ctagaggtgg atctggagct gcagctggat ctggttctgg ttctatgcgt aaaggcgaag
     2821 agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
     2881 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
     2941 tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt
     3001 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
     3061 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
     3121 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
     3181 aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa tacaatttta
     3241 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     3301 aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     3361 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     3421 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     3481 cagcgggcat cacgcatggt atggatgaac tgtacaaagg cagcagccat catcatcatc
     3541 atcaccactg aaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg
     3601 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3661 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3721 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3781 ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag cctagaagat
     3841 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     3901 ctatcataac ccctagggta tacccatcta attggaacca gataagtgaa atctagttcc
     3961 aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc
     4021 tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc
     4081 caactatttt gtccgcccac aaccggttga cttgggtcaa ctgtcagacc aagtttactc
     4141 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4201 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4441 tctagtgtag ccgtagttag gccaccactt caagaact
//
