LOCUS       sequence-298332-        6183 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 6183)
  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 6183)
  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-frag1_nostop
FEATURES             Location/Qualifiers
     source          1..6183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     414..431
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(590..617)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(709..795)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     844..866
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     858..875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     859..875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    891..990
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4630..4729)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4746..4765)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4747..4765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4770..4786)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4786)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4888..4907)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4899..5708
                     /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(4966..4985)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(5855..6183,1..260)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
       61 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      301 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      361 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      421 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      481 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      541 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
      601 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
      661 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
      721 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
      781 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
      841 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga
      901 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta
      961 taatgccaac tttgtacaaa aaagttggca tgttcgtgtt cctggtgctg ctgccactgg
     1021 tgagctccca gtgcgtgaac ctgaccacca gaacccagct gccacctgcc tacaccaact
     1081 ccttcaccag gggcgtgtac tacccagaca aggtgttcag atccagcgtg ctgcactcta
     1141 cccaggacct gttcctgcca ttcttctcta acgtgacctg gttccacgcc atccacgtgt
     1201 ctggaaccaa cggcaccaag agattcgaca acccagtgct gccattcaac gacggcgtgt
     1261 acttcgcttc taccgagaag tctaacatca tcaggggttg gatcttcgga accaccctgg
     1321 actctaagac ccagtctctg ctgatcgtga acaacgctac caacgtggtg atcaaggtgt
     1381 gcgagttcca gttctgcaac gacccattcc tgggagtgta ctaccacaag aacaacaagt
     1441 cttggatgga aagcgagttc agggtgtaca gctctgctaa caactgcacc ttcgagtacg
     1501 tgtctcagcc tttcctgatg gacctggaag gaaagcaggg aaacttcaag aacctgagag
     1561 agttcgtgtt caagaacatc gacggttact tcaagatata cagcaagcac accccaatca
     1621 acctggtgag ggacctgcca cagggtttct ctgctctgga gcccctggtg gacctgccaa
     1681 tcggcatcaa catcaccagg ttccagaccc tgctggctct gcacaggtct tacctgaccc
     1741 caggcgacag cagctctgga tggaccgccg gcgccgctgc ttactacgtg ggatacctgc
     1801 agccaaggac cttcctgctg aagtacaacg aaaacggcac catcaccgac gccgtggact
     1861 gcgctctgga cccactgtct gagaccaagt gcaccctgaa gtctttcacc gtggaaaagg
     1921 gaatctacca gaccagcaac ttccgcgtgc agcccaccga gtctatcgtg agattcccaa
     1981 acatcaccaa cctgtgccca ttcggagaag tgttcaacgc taccaggttc gccagcgtgt
     2041 acgcttggaa caggaagaga atcagcaact gcgtggccga ctactctgtg ctgtacaact
     2101 ctgcctcctt ctccaccttc aagtgctacg gcgtgtcccc aaccaagctg aacgacctgt
     2161 gcttcaccaa cgtgtacgct gactccttcg tgatcagagg agacgaagtg agacagatcg
     2221 ctcccggtca gaccggaaag atcgccgact acaactacaa gctgccagac gacttcaccg
     2281 gttgcgtgat cgcttggaac tctaacaacc tggactctaa ggtgggaggc aactacaact
     2341 acctgtacag gctgttccgc aagagcaacc tgaagccttt cgaaagggac atcagcaccg
     2401 aaatctacca ggctggatct accccatgca acggagtgga aggattcaac tgctacttcc
     2461 ctctgcagtc ttacggattc cagccaacca acggagtggg ataccagcca tacagagtgg
     2521 tggtgctgtc tttcgagctg ctgcacgctc ctgctaccgt gtgcggacca aagaagtcta
     2581 ccaacctggt gaagaacaag tgcgtgaact tcaacttcaa cggactgacc ggaaccggag
     2641 tgctgaccga atccaacaag aagttcctgc ctttccagca gttcggtaga gacatcgccg
     2701 acaccaccga cgctgtgaga gacccacaga ccctggaaat cctggacatc accccctgct
     2761 ctttcggagg agtgagcgtg atcaccccag gaaccaacac ctccaaccag gtggccgtgc
     2821 tgtaccagga cgtgaactgc accgaagtgc cagtggccat ccacgctgac cagctgaccc
     2881 caacctggag agtgtacagc accggatcta acgtgttcca gacccgtgcc ggctgcctga
     2941 tcggagccga gcacgtgaac aactcctacg aatgcgacat cccaatcgga gctggcatct
     3001 gcgcttctta ccagacccag accaacagcc caagaagggc cagatccgtg gcctctcaga
     3061 gcatcatcgc ctacaccatg agcctgggcg ctgaaaacag cgtggcttac agcaacaact
     3121 ctatcgccat cccaaccaac ttcaccatct ctgtgaccac cgaaatcctg ccagtgagca
     3181 tgaccaagac cagcgtggac tgcaccatgt acatctgcgg agactctacc gaatgctcta
     3241 acctgctgct gcagtacggc agcttctgca cccagctgaa ccgcgctctg accggaatcg
     3301 ctgtggagca ggacaagaac acccaggaag tgttcgctca ggtgaagcag atatacaaga
     3361 ccccacctat caaggacttc ggaggattca acttctctca gatcctgccc gacccatcta
     3421 agccctctaa gaggtctttc atcgaagacc tgctgttcaa caaggtgacc ctggccgacg
     3481 ccggtttcat caagcagtac ggtgactgcc tgggagacat cgccgctaga gacctgatct
     3541 gcgcccagaa gttcaacggt ctgaccgtgc tgccaccact gctgaccgac gagatgatcg
     3601 cccagtacac ctctgccctg ctggctggaa ccatcacctc tggttggacc ttcggagctg
     3661 gcgccgccct gcagatccca ttcgctatgc agatggccta ccgtttcaac ggtatcggag
     3721 tgacccagaa cgtgctgtac gaaaaccaga agctgatcgc taaccagttc aactctgcta
     3781 tcggaaagat ccaggactcc ctgtcctcta ccgcttctgc cctgggaaag ctgcaggacg
     3841 tggtgaacca gaacgcccag gccctgaaca ccctggtgaa gcagctgtct tctaacttcg
     3901 gagccatctc ctctgtgctg aacgacatcc tgtctaggct ggacaaggtg gaagccgagg
     3961 tgcagatcga cagactgatc accggaagac tgcagtctct gcagacctac gtgacccagc
     4021 agctgatcag ggccgctgaa atcagagctt ctgccaacct ggctgccacc aagatgtccg
     4081 aatgcgtgct gggccagtct aagagggtgg acttctgcgg aaagggatac cacctgatgt
     4141 ccttcccaca gtctgctcca cacggagtgg tgttcctgca cgtgacctac gtgcccgccc
     4201 aggaaaagaa cttcaccacc gccccagcta tctgccacga cggaaaggct cacttcccca
     4261 gggagggagt gttcgtgtct aacggaaccc actggttcgt gacccagagg aacttctacg
     4321 agccacagat catcaccacc gacaacacct tcgtgtctgg aaactgcgac gtggtgatcg
     4381 gaatcgtgaa caacaccgtg tacgacccac tgcagccaga actggactct ttcaaggagg
     4441 aactggacaa gtacttcaag aaccacacct ctccagacgt ggacctggga gacatctccg
     4501 gaatcaacgc ctctgtggtg aacatccaga aggaaatcga cagactgaac gaggtggcca
     4561 agaacctgaa cgaatctctg atcgacctgc aggaactggg aaagtacgag cagtacatca
     4621 agtggcccta cccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     4681 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     4741 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     4801 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     4861 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     4921 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4981 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     5041 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     5101 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     5161 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     5221 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     5281 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     5341 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     5401 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     5461 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     5521 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     5581 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     5641 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     5701 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     5761 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     5821 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5881 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5941 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6001 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6061 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6121 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6181 gtt
//
