LOCUS       sequence-298337-        6141 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 6141)
  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 6141)
  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-frag2_nostop
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(147..174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(266..352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     401..423
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     415..432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     416..432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    448..547
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4145..4244)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4261..4280)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4262..4280)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4285..4301)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4285..4301)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4403..4422)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4414..5223
                     /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(4481..4500)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5370..5958
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5859..5878
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6112..6129
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
      241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
      301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      361 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      421 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      481 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      541 gttggcatgg tgaacctgac caccagaacc cagctgccac ctgcctacac caactccttc
      601 accaggggcg tgtactaccc agacaaggtg ttcagatcca gcgtgctgca ctctacccag
      661 gacctgttcc tgccattctt ctctaacgtg acctggttcc acgccatcca cgtgtctgga
      721 accaacggca ccaagagatt cgacaaccca gtgctgccat tcaacgacgg cgtgtacttc
      781 gcttctaccg agaagtctaa catcatcagg ggttggatct tcggaaccac cctggactct
      841 aagacccagt ctctgctgat cgtgaacaac gctaccaacg tggtgatcaa ggtgtgcgag
      901 ttccagttct gcaacgaccc attcctggga gtgtactacc acaagaacaa caagtcttgg
      961 atggaaagcg agttcagggt gtacagctct gctaacaact gcaccttcga gtacgtgtct
     1021 cagcctttcc tgatggacct ggaaggaaag cagggaaact tcaagaacct gagagagttc
     1081 gtgttcaaga acatcgacgg ttacttcaag atatacagca agcacacccc aatcaacctg
     1141 gtgagggacc tgccacaggg tttctctgct ctggagcccc tggtggacct gccaatcggc
     1201 atcaacatca ccaggttcca gaccctgctg gctctgcaca ggtcttacct gaccccaggc
     1261 gacagcagct ctggatggac cgccggcgcc gctgcttact acgtgggata cctgcagcca
     1321 aggaccttcc tgctgaagta caacgaaaac ggcaccatca ccgacgccgt ggactgcgct
     1381 ctggacccac tgtctgagac caagtgcacc ctgaagtctt tcaccgtgga aaagggaatc
     1441 taccagacca gcaacttccg cgtgcagccc accgagtcta tcgtgagatt cccaaacatc
     1501 accaacctgt gcccattcgg agaagtgttc aacgctacca ggttcgccag cgtgtacgct
     1561 tggaacagga agagaatcag caactgcgtg gccgactact ctgtgctgta caactctgcc
     1621 tccttctcca ccttcaagtg ctacggcgtg tccccaacca agctgaacga cctgtgcttc
     1681 accaacgtgt acgctgactc cttcgtgatc agaggagacg aagtgagaca gatcgctccc
     1741 ggtcagaccg gaaagatcgc cgactacaac tacaagctgc cagacgactt caccggttgc
     1801 gtgatcgctt ggaactctaa caacctggac tctaaggtgg gaggcaacta caactacctg
     1861 tacaggctgt tccgcaagag caacctgaag cctttcgaaa gggacatcag caccgaaatc
     1921 taccaggctg gatctacccc atgcaacgga gtggaaggat tcaactgcta cttccctctg
     1981 cagtcttacg gattccagcc aaccaacgga gtgggatacc agccatacag agtggtggtg
     2041 ctgtctttcg agctgctgca cgctcctgct accgtgtgcg gaccaaagaa gtctaccaac
     2101 ctggtgaaga acaagtgcgt gaacttcaac ttcaacggac tgaccggaac cggagtgctg
     2161 accgaatcca acaagaagtt cctgcctttc cagcagttcg gtagagacat cgccgacacc
     2221 accgacgctg tgagagaccc acagaccctg gaaatcctgg acatcacccc ctgctctttc
     2281 ggaggagtga gcgtgatcac cccaggaacc aacacctcca accaggtggc cgtgctgtac
     2341 caggacgtga actgcaccga agtgccagtg gccatccacg ctgaccagct gaccccaacc
     2401 tggagagtgt acagcaccgg atctaacgtg ttccagaccc gtgccggctg cctgatcgga
     2461 gccgagcacg tgaacaactc ctacgaatgc gacatcccaa tcggagctgg catctgcgct
     2521 tcttaccaga cccagaccaa cagcccaaga agggccagat ccgtggcctc tcagagcatc
     2581 atcgcctaca ccatgagcct gggcgctgaa aacagcgtgg cttacagcaa caactctatc
     2641 gccatcccaa ccaacttcac catctctgtg accaccgaaa tcctgccagt gagcatgacc
     2701 aagaccagcg tggactgcac catgtacatc tgcggagact ctaccgaatg ctctaacctg
     2761 ctgctgcagt acggcagctt ctgcacccag ctgaaccgcg ctctgaccgg aatcgctgtg
     2821 gagcaggaca agaacaccca ggaagtgttc gctcaggtga agcagatata caagacccca
     2881 cctatcaagg acttcggagg attcaacttc tctcagatcc tgcccgaccc atctaagccc
     2941 tctaagaggt ctttcatcga agacctgctg ttcaacaagg tgaccctggc cgacgccggt
     3001 ttcatcaagc agtacggtga ctgcctggga gacatcgccg ctagagacct gatctgcgcc
     3061 cagaagttca acggtctgac cgtgctgcca ccactgctga ccgacgagat gatcgcccag
     3121 tacacctctg ccctgctggc tggaaccatc acctctggtt ggaccttcgg agctggcgcc
     3181 gccctgcaga tcccattcgc tatgcagatg gcctaccgtt tcaacggtat cggagtgacc
     3241 cagaacgtgc tgtacgaaaa ccagaagctg atcgctaacc agttcaactc tgctatcgga
     3301 aagatccagg actccctgtc ctctaccgct tctgccctgg gaaagctgca ggacgtggtg
     3361 aaccagaacg cccaggccct gaacaccctg gtgaagcagc tgtcttctaa cttcggagcc
     3421 atctcctctg tgctgaacga catcctgtct aggctggaca aggtggaagc cgaggtgcag
     3481 atcgacagac tgatcaccgg aagactgcag tctctgcaga cctacgtgac ccagcagctg
     3541 atcagggccg ctgaaatcag agcttctgcc aacctggctg ccaccaagat gtccgaatgc
     3601 gtgctgggcc agtctaagag ggtggacttc tgcggaaagg gataccacct gatgtccttc
     3661 ccacagtctg ctccacacgg agtggtgttc ctgcacgtga cctacgtgcc cgcccaggaa
     3721 aagaacttca ccaccgcccc agctatctgc cacgacggaa aggctcactt ccccagggag
     3781 ggagtgttcg tgtctaacgg aacccactgg ttcgtgaccc agaggaactt ctacgagcca
     3841 cagatcatca ccaccgacaa caccttcgtg tctggaaact gcgacgtggt gatcggaatc
     3901 gtgaacaaca ccgtgtacga cccactgcag ccagaactgg actctttcaa ggaggaactg
     3961 gacaagtact tcaagaacca cacctctcca gacgtggacc tgggagacat ctccggaatc
     4021 aacgcctctg tggtgaacat ccagaaggaa atcgacagac tgaacgaggt ggccaagaac
     4081 ctgaacgaat ctctgatcga cctgcaggaa ctgggaaagt acgagcagta catcaagtgg
     4141 ccctacccaa ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     4201 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     4261 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     4321 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     4381 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg
     4441 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     4501 aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag
     4561 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     4621 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4681 gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa
     4741 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4801 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4861 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4921 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4981 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     5041 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     5101 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     5161 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     5221 taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac
     5281 ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5401 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5461 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5521 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5581 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5641 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5701 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5761 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5821 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5881 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5941 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6001 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6061 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6121 gtgagcgagg aagcggaaga g
//
