LOCUS       sequence-298348-        4128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with a fragmented insert of S 
            gene's CDS from SARS-CoV-2 isolate Wuhan-Hu-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  AUTHORS   Kim DK, Knapp JJ, Kuang D, Chawla A, Cassonnet P, Lee H, 
            Sheykhkarimli D, Samavarchi-Tehrani P, Abdouni H, Rayhan A, Li R, 
            Pogoutse O, Coyaud E, van der Werf S, Demeret C, Gingras AC, Taipale
            M, Raught B, Jacob Y, Roth FP
  TITLE     A Comprehensive, Flexible Collection of SARS-CoV-2 Coding Regions.
  JOURNAL   G3 (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554.
  PUBMED    32763951
REFERENCE   2  (bases 1 to 4128)
  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: "G3 
            (Bethesda). 2020 Aug 6. pii: g3.120.401554. doi: 
            10.1534/g3.120.401554."
COMMENT     Sequence Label: pDONR221 SARS-CoV-2 S-frag5_nostop
FEATURES             Location/Qualifiers
     source          1..4128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..46
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     38..55
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     39..55
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    71..170
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1755..1854)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1871..1890)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1872..1890)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1895..1911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1895..1911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2013..2032)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2024..2833
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2091..2110)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2980..3568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3469..3488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3722..3739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3898..3925)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4017..4103)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
       61 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
      121 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagttggca tggtggcctc
      181 tcagagcatc atcgcctaca ccatgagcct gggcgctgaa aacagcgtgg cttacagcaa
      241 caactctatc gccatcccaa ccaacttcac catctctgtg accaccgaaa tcctgccagt
      301 gagcatgacc aagaccagcg tggactgcac catgtacatc tgcggagact ctaccgaatg
      361 ctctaacctg ctgctgcagt acggcagctt ctgcacccag ctgaaccgcg ctctgaccgg
      421 aatcgctgtg gagcaggaca agaacaccca ggaagtgttc gctcaggtga agcagatata
      481 caagacccca cctatcaagg acttcggagg attcaacttc tctcagatcc tgcccgaccc
      541 atctaagccc tctaagaggt ctttcatcga agacctgctg ttcaacaagg tgaccctggc
      601 cgacgccggt ttcatcaagc agtacggtga ctgcctggga gacatcgccg ctagagacct
      661 gatctgcgcc cagaagttca acggtctgac cgtgctgcca ccactgctga ccgacgagat
      721 gatcgcccag tacacctctg ccctgctggc tggaaccatc acctctggtt ggaccttcgg
      781 agctggcgcc gccctgcaga tcccattcgc tatgcagatg gcctaccgtt tcaacggtat
      841 cggagtgacc cagaacgtgc tgtacgaaaa ccagaagctg atcgctaacc agttcaactc
      901 tgctatcgga aagatccagg actccctgtc ctctaccgct tctgccctgg gaaagctgca
      961 ggacgtggtg aaccagaacg cccaggccct gaacaccctg gtgaagcagc tgtcttctaa
     1021 cttcggagcc atctcctctg tgctgaacga catcctgtct aggctggaca aggtggaagc
     1081 cgaggtgcag atcgacagac tgatcaccgg aagactgcag tctctgcaga cctacgtgac
     1141 ccagcagctg atcagggccg ctgaaatcag agcttctgcc aacctggctg ccaccaagat
     1201 gtccgaatgc gtgctgggcc agtctaagag ggtggacttc tgcggaaagg gataccacct
     1261 gatgtccttc ccacagtctg ctccacacgg agtggtgttc ctgcacgtga cctacgtgcc
     1321 cgcccaggaa aagaacttca ccaccgcccc agctatctgc cacgacggaa aggctcactt
     1381 ccccagggag ggagtgttcg tgtctaacgg aacccactgg ttcgtgaccc agaggaactt
     1441 ctacgagcca cagatcatca ccaccgacaa caccttcgtg tctggaaact gcgacgtggt
     1501 gatcggaatc gtgaacaaca ccgtgtacga cccactgcag ccagaactgg actctttcaa
     1561 ggaggaactg gacaagtact tcaagaacca cacctctcca gacgtggacc tgggagacat
     1621 ctccggaatc aacgcctctg tggtgaacat ccagaaggaa atcgacagac tgaacgaggt
     1681 ggccaagaac ctgaacgaat ctctgatcga cctgcaggaa ctgggaaagt acgagcagta
     1741 catcaagtgg ccctacccaa ctttcttgta caaagttggc attataagaa agcattgctt
     1801 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1861 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1921 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1981 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2041 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2101 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2161 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2221 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2281 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2341 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2401 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2461 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2521 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2581 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2641 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2701 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2761 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2821 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2881 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3721 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3781 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3841 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3901 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     3961 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     4021 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     4081 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgc
//
