LOCUS       sequence-298343-        4560 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 4560)
  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 4560)
  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-frag4_nostop
FEATURES             Location/Qualifiers
     source          1..4560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(47..74)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(166..252)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     301..323
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     315..332
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     316..332
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    348..447
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2464..2563)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2580..2599)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2581..2599)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2604..2620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2604..2620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2722..2741)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2733..3542
                     /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(2800..2819)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3689..4277
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4178..4197
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4431..4448
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
       61 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
      121 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
      181 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
      241 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
      301 cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt
      361 tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa
      421 tgccaacttt gtacaaaaaa gttggcatgg tgaacctgac caccagaacc cagctgccac
      481 ctgcctacac caactccttc accaggggcg tgtactaccc agacaaggtg ttcagatcca
      541 gcgtgctgca ctctacccag gacctgttcc tgccattctt ctctaacgtg acctggttcc
      601 acgccatcca cgtgtctgga accaacggca ccaagagatt cgacaaccca gtgctgccat
      661 tcaacgacgg cgtgtacttc gcttctaccg agaagtctaa catcatcagg ggttggatct
      721 tcggaaccac cctggactct aagacccagt ctctgctgat cgtgaacaac gctaccaacg
      781 tggtgatcaa ggtgtgcgag ttccagttct gcaacgaccc attcctggga gtgtactacc
      841 acaagaacaa caagtcttgg atggaaagcg agttcagggt gtacagctct gctaacaact
      901 gcaccttcga gtacgtgtct cagcctttcc tgatggacct ggaaggaaag cagggaaact
      961 tcaagaacct gagagagttc gtgttcaaga acatcgacgg ttacttcaag atatacagca
     1021 agcacacccc aatcaacctg gtgagggacc tgccacaggg tttctctgct ctggagcccc
     1081 tggtggacct gccaatcggc atcaacatca ccaggttcca gaccctgctg gctctgcaca
     1141 ggtcttacct gaccccaggc gacagcagct ctggatggac cgccggcgcc gctgcttact
     1201 acgtgggata cctgcagcca aggaccttcc tgctgaagta caacgaaaac ggcaccatca
     1261 ccgacgccgt ggactgcgct ctggacccac tgtctgagac caagtgcacc ctgaagtctt
     1321 tcaccgtgga aaagggaatc taccagacca gcaacttccg cgtgcagccc accgagtcta
     1381 tcgtgagatt cccaaacatc accaacctgt gcccattcgg agaagtgttc aacgctacca
     1441 ggttcgccag cgtgtacgct tggaacagga agagaatcag caactgcgtg gccgactact
     1501 ctgtgctgta caactctgcc tccttctcca ccttcaagtg ctacggcgtg tccccaacca
     1561 agctgaacga cctgtgcttc accaacgtgt acgctgactc cttcgtgatc agaggagacg
     1621 aagtgagaca gatcgctccc ggtcagaccg gaaagatcgc cgactacaac tacaagctgc
     1681 cagacgactt caccggttgc gtgatcgctt ggaactctaa caacctggac tctaaggtgg
     1741 gaggcaacta caactacctg tacaggctgt tccgcaagag caacctgaag cctttcgaaa
     1801 gggacatcag caccgaaatc taccaggctg gatctacccc atgcaacgga gtggaaggat
     1861 tcaactgcta cttccctctg cagtcttacg gattccagcc aaccaacgga gtgggatacc
     1921 agccatacag agtggtggtg ctgtctttcg agctgctgca cgctcctgct accgtgtgcg
     1981 gaccaaagaa gtctaccaac ctggtgaaga acaagtgcgt gaacttcaac ttcaacggac
     2041 tgaccggaac cggagtgctg accgaatcca acaagaagtt cctgcctttc cagcagttcg
     2101 gtagagacat cgccgacacc accgacgctg tgagagaccc acagaccctg gaaatcctgg
     2161 acatcacccc ctgctctttc ggaggagtga gcgtgatcac cccaggaacc aacacctcca
     2221 accaggtggc cgtgctgtac caggacgtga actgcaccga agtgccagtg gccatccacg
     2281 ctgaccagct gaccccaacc tggagagtgt acagcaccgg atctaacgtg ttccagaccc
     2341 gtgccggctg cctgatcgga gccgagcacg tgaacaactc ctacgaatgc gacatcccaa
     2401 tcggagctgg catctgcgct tcttaccaga cccagaccaa cagcccaaga agggccagat
     2461 cctacccaac tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2521 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     2581 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2641 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2701 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2761 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2821 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2881 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     2941 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     3001 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     3061 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3121 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     3181 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3241 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3301 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3361 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3421 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3481 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3541 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3601 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3661 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3721 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3781 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3841 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3901 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3961 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4021 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4081 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4141 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4201 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4261 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4321 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4381 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4441 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4501 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
//
