LOCUS       sequence-298338-        4602 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 4602)
  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 4602)
  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-frag3_nostop
FEATURES             Location/Qualifiers
     source          1..4602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(165..192)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(284..370)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     419..441
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     433..450
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     434..450
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    466..565
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2624..2723)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2740..2759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2741..2759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2764..2780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2764..2780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2882..2901)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2893..3702
                     /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(2960..2979)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3849..4437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4338..4357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(4591..4602,1..6)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
      181 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
      241 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
      301 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
      361 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
      421 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
      481 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg
      541 ccaactttgt acaaaaaagt tggcatgttc gtgttcctgg tgctgctgcc actggtgagc
      601 tcccagtgcg tgaacctgac caccagaacc cagctgccac ctgcctacac caactccttc
      661 accaggggcg tgtactaccc agacaaggtg ttcagatcca gcgtgctgca ctctacccag
      721 gacctgttcc tgccattctt ctctaacgtg acctggttcc acgccatcca cgtgtctgga
      781 accaacggca ccaagagatt cgacaaccca gtgctgccat tcaacgacgg cgtgtacttc
      841 gcttctaccg agaagtctaa catcatcagg ggttggatct tcggaaccac cctggactct
      901 aagacccagt ctctgctgat cgtgaacaac gctaccaacg tggtgatcaa ggtgtgcgag
      961 ttccagttct gcaacgaccc attcctggga gtgtactacc acaagaacaa caagtcttgg
     1021 atggaaagcg agttcagggt gtacagctct gctaacaact gcaccttcga gtacgtgtct
     1081 cagcctttcc tgatggacct ggaaggaaag cagggaaact tcaagaacct gagagagttc
     1141 gtgttcaaga acatcgacgg ttacttcaag atatacagca agcacacccc aatcaacctg
     1201 gtgagggacc tgccacaggg tttctctgct ctggagcccc tggtggacct gccaatcggc
     1261 atcaacatca ccaggttcca gaccctgctg gctctgcaca ggtcttacct gaccccaggc
     1321 gacagcagct ctggatggac cgccggcgcc gctgcttact acgtgggata cctgcagcca
     1381 aggaccttcc tgctgaagta caacgaaaac ggcaccatca ccgacgccgt ggactgcgct
     1441 ctggacccac tgtctgagac caagtgcacc ctgaagtctt tcaccgtgga aaagggaatc
     1501 taccagacca gcaacttccg cgtgcagccc accgagtcta tcgtgagatt cccaaacatc
     1561 accaacctgt gcccattcgg agaagtgttc aacgctacca ggttcgccag cgtgtacgct
     1621 tggaacagga agagaatcag caactgcgtg gccgactact ctgtgctgta caactctgcc
     1681 tccttctcca ccttcaagtg ctacggcgtg tccccaacca agctgaacga cctgtgcttc
     1741 accaacgtgt acgctgactc cttcgtgatc agaggagacg aagtgagaca gatcgctccc
     1801 ggtcagaccg gaaagatcgc cgactacaac tacaagctgc cagacgactt caccggttgc
     1861 gtgatcgctt ggaactctaa caacctggac tctaaggtgg gaggcaacta caactacctg
     1921 tacaggctgt tccgcaagag caacctgaag cctttcgaaa gggacatcag caccgaaatc
     1981 taccaggctg gatctacccc atgcaacgga gtggaaggat tcaactgcta cttccctctg
     2041 cagtcttacg gattccagcc aaccaacgga gtgggatacc agccatacag agtggtggtg
     2101 ctgtctttcg agctgctgca cgctcctgct accgtgtgcg gaccaaagaa gtctaccaac
     2161 ctggtgaaga acaagtgcgt gaacttcaac ttcaacggac tgaccggaac cggagtgctg
     2221 accgaatcca acaagaagtt cctgcctttc cagcagttcg gtagagacat cgccgacacc
     2281 accgacgctg tgagagaccc acagaccctg gaaatcctgg acatcacccc ctgctctttc
     2341 ggaggagtga gcgtgatcac cccaggaacc aacacctcca accaggtggc cgtgctgtac
     2401 caggacgtga actgcaccga agtgccagtg gccatccacg ctgaccagct gaccccaacc
     2461 tggagagtgt acagcaccgg atctaacgtg ttccagaccc gtgccggctg cctgatcgga
     2521 gccgagcacg tgaacaactc ctacgaatgc gacatcccaa tcggagctgg catctgcgct
     2581 tcttaccaga cccagaccaa cagcccaaga agggccagat cctacccaac tttcttgtac
     2641 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     2701 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     2761 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     2821 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2881 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     2941 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     3001 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     3061 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     3121 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     3181 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3241 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     3301 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3361 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3421 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3481 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     3541 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     3601 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     3661 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     3721 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     3781 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3841 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3901 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3961 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4021 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4081 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4141 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4201 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4261 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4321 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4381 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4441 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4501 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4561 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tg
//
