LOCUS       sequence-304666-        4317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4317)
  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 4317)
  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: pDONR223 SARS-CoV2 TEV-NSP4
FEATURES             Location/Qualifiers
     source          1..4317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(83..110)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(202..288)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     337..359
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     351..368
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     352..368
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    384..483
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             483..503
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(2011..2110)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2127..2146)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2128..2146)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2151..2167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2151..2167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2598..3389
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3482..4070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3971..3990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4224..4241
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
       61 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
      121 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
      181 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      241 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
      301 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
      361 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      421 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      481 gcgaaaacct gtacttccaa ggcatgaaga tcgtgaacaa ctggctgaag cagctgatca
      541 aggtgaccct ggtgttcctg ttcgtggccg ctatcttcta cctgatcacc cctgtgcacg
      601 tgatgagcaa gcacaccgac ttcagctccg agatcatcgg ctacaaggcc atcgacggcg
      661 gagtgaccag agacatcgct tccaccgaca cctgcttcgc caacaagcac gctgacttcg
      721 acacctggtt ctcccagaga ggtggctctt acaccaacga caaggcctgc ccactgatcg
      781 ccgctgtgat cacccgtgaa gtgggattcg tggtgcctgg cctgccagga accatcctga
      841 gaaccaccaa cggtgacttc ctgcacttcc tgccacgcgt gttcagcgcc gtgggcaaca
      901 tctgctacac cccttctaag ctgatcgagt acaccgactt cgccacctct gcttgcgtgc
      961 tggccgctga atgcaccatc ttcaaggacg ccagcggaaa gccagtgccc tactgctacg
     1021 acaccaacgt gctggagggt agcgtggctt acgaatccct gcgcccagac accagatacg
     1081 tgctgatgga cggctccatc atccagttcc ccaacaccta cctggaggga agcgtgcgcg
     1141 tggtgaccac cttcgactcc gaatactgca gacacggaac ctgcgagcgt tccgaagccg
     1201 gagtgtgcgt gtctaccagc ggtcgctggg tgctgaacaa cgactactac aggtctctgc
     1261 ctggcgtgtt ctgcggagtg gatgctgtga acctgctgac caacatgttc accccactga
     1321 tccagcccat cggtgctctg gacatctccg cctctatcgt ggctggaggt atcgtggcca
     1381 tcgtggtgac ctgcctggct tactacttca tgaggttccg cagggccttc ggtgaataca
     1441 gccacgtggt ggctttcaac accctgctgt tcctgatgtc cttcaccgtg ctgtgcctga
     1501 cccccgtgta ctctttcctg ccaggcgtgt acagcgtgat ctacctgtac ctgaccttct
     1561 acctgaccaa cgacgtgtct ttcctggccc acatccagtg gatggtcatg ttcacccctc
     1621 tggtgccttt ctggatcacc atcgcttaca tcatctgcat ctctaccaag cacttctact
     1681 ggttcttcag caactacctg aagagacgtg tggtgttcaa cggcgtgagc ttctccacct
     1741 tcgaggaagc cgctctgtgc accttcctgc tgaacaagga gatgtacctg aagctgcgca
     1801 gcgacgtgct gctgcctctg acccagtaca acaggtacct ggccctgtac aacaagtaca
     1861 agtacttctc cggcgctatg gacaccacct cttaccgtga agccgcttgc tgccacctgg
     1921 ccaaggctct gaacgacttc tctaacagcg gatccgacgt gctgtaccag ccccctcaga
     1981 cctctatcac cagcgccgtg ctgcagtaat acccaacttt cttgtacaaa gttggcatta
     2041 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     2101 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     2161 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     2221 atatatcatc atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
     2281 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     2341 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     2401 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     2461 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     2521 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     2581 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     2641 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     2701 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     2761 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     2821 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     2881 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     2941 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     3001 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     3061 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     3121 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     3181 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     3241 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     3301 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     3361 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     3421 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3481 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3541 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3601 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3661 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3721 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3781 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3841 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3901 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3961 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4021 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4081 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4141 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4201 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4261 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgaca
//
