LOCUS       sequence-307796-        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      190..778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     679..698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  913..1136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1203..1736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1925..1930,1931..1947,1948..1953))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2134..2225
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2150..2167
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2993..3016
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3062..3775
                     /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             3785..3805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3938..4072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3975..3994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4029..4048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4193..4358
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcgctcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     1741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     1801 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     1861 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     1921 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     1981 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gccataactt cgtatagcat
     2041 acattatacg aagttatctg taactataac ggtcctaagg tagcgagttt aaacactagt
     2101 atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa aatgataacc
     2161 atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta
     2221 taaatattcc ggattattca taccgtccca ccatcgggcg cggatccctc gagccaccat
     2281 gccattcggt tggttgatag ttggtgttgc gttgcttgca gtctttcagt cagcttctaa
     2341 gataatcact ctgaagaagc gttggcaact ggcgctctca aaaggcgttc acttcgtgtg
     2401 caacctcctg ttgctctttg tcaccgttta ctcacatctc ttgcttgttg ctgctggcct
     2461 ggaagctccc ttcctttacc tgtacgctct ggtctatttt ctgcagagca taaattttgt
     2521 caggataata atgagactgt ggctttgttg gaaatgccgc tccaagaacc cgcttttgta
     2581 cgatgccaat tacttcctgt gttggcatac gaattgctac gactactgta ttccgtataa
     2641 tagcgtcact tcttcaatcg tgataacgtc cggtgacggt acaacatcac ctatttctga
     2701 acacgattac caaataggtg gctacaccga gaaatgggag agtggagtca aagattgtgt
     2761 agtgctgcac tcatatttta cctcggatta ttatcaactc tactcaacac agttgtctac
     2821 tgatacagga gttgagcacg taacattttt catttacaat aaaattgtgg acgaacccga
     2881 ggagcatgta cagattcata ctatcgacgg tagcagcggt gtcgtgaacc ccgttatgga
     2941 gccaatttac gacgaaccca ccacgacaac atccgtcccc ctgtcgaatt ctttggaagt
     3001 tttgtttcaa ggtccgtcta gaggtggatc tggagctgca gctggatctg gttctggttc
     3061 tatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
     3121 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
     3181 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt ggccgactct
     3241 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
     3301 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
     3361 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
     3421 aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc tgggccataa
     3481 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     3541 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     3601 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     3661 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     3721 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaaggcag
     3781 cagccatcat catcatcatc accactgaaa gcttgtcgag aagtactaga ggatcataat
     3841 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3901 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3961 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4021 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     4081 gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     4141 ccaccgctga gcaataacta tcataacccc tagggtatac ccatctaatt ggaaccagat
     4201 aagtgaaatc tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc
     4261 tacacccagt tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt
     4321 ccacccctcc cagttcccaa ctattttgtc cgcccaca
//
