LOCUS       sequence-333052-        4296 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 4296)
  AUTHORS   Weller B, Lin CW, Pogoutse O, Sauer M, Marin-de la Rosa N, Strobel 
            A, Young V, Knapp JJ, Rayhan A, Falter C, Kim DK, Roth FP, 
            Falter-Braun P
  TITLE     A resource of human coronavirus protein-coding sequences in a 
            flexible, multipurpose Gateway Entry clone collection.
  JOURNAL   G3 (Bethesda). 2023 Jul 5;13(7):jkad105. doi: 
            10.1093/g3journal/jkad105.
  PUBMED    37267226
REFERENCE   2  (bases 1 to 4296)
  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)."; date: "2023-07-5"; volume: "13(7):jkad105. doi"; 
            pages: " 10.1093/g3journal/jkad105"
COMMENT     Sequence Label: pDONR223 SARS-CoV-1 NSP4_stop
FEATURES             Location/Qualifiers
     source          1..4296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1639..1738)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1755..1774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1756..1774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1779..1795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1779..1795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2226..3017
                     /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      3110..3698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3599..3618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3852..3869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4028..4055)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4147..4233)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4282..4296,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4296,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catgaagatt gttagtactt gttttaaact tatgcttaag gccacattat
      181 tgtgcgttct tgctgcattg gtttgttata tcgttatgcc agtacataca ttgtcaatcc
      241 atgatggtta cacaaatgaa atcattggtt acaaagccat tcaggatggt gtcactcgtg
      301 acatcatttc tactgatgat tgttttgcaa ataaacatgc tggttttgac gcatggttta
      361 gccagcgtgg tggttcatac aaaaatgaca aaagctgccc tgtagtagct gctatcatta
      421 caagagagat tggtttcata gtgcctggct taccgggtac tgtgctgaga gcaatcaatg
      481 gtgacttctt gcattttcta cctcgtgttt ttagtgctgt tggcaacatt tgctacacac
      541 cttccaaact cattgagtat agtgattttg ctacctctgc ttgcgttctt gctgctgagt
      601 gtacaatttt taaggatgct atgggcaaac ctgtgccata ttgttatgac actaatttgc
      661 tagagggttc tatttcttat agtgagcttc gtccagacac tcgttatgtg cttatggatg
      721 gttccatcat acagtttcct aacacttacc tggagggttc tgttagagta gtaacaactt
      781 ttgatgctga gtactgtaga catggtacat gcgaaaggtc agaagtaggt atttgcctat
      841 ctaccagtgg tagatgggtt cttaataatg agcattacag agctctatca ggagttttct
      901 gtggtgttga tgcgatgaat ctcatagcta acatctttac tcctcttgtg caacctgtgg
      961 gtgctttaga tgtgtctgct tcagtagtgg ctggtggtat tattgccata ttggtgactt
     1021 gtgctgccta ctactttatg aaattcagac gtgtttttgg tgagtacaac catgttgttg
     1081 ctgctaatgc acttttgttt ttgatgtctt tcactatact ctgtctggta ccagcttaca
     1141 gctttctgcc gggagtctac tcagtctttt acttgtactt gacattctat ttcaccaatg
     1201 atgtttcatt cttggctcac cttcaatggt ttgccatgtt ttctcctatt gtgccttttt
     1261 ggataacagc aatctatgta ttctgtattt ctctgaagca ctgccattgg ttctttaaca
     1321 actatcttag gaaaagagtc atgtttaatg gagttacatt tagtaccttc gaggaggctg
     1381 ctttgtgtac ctttttgctc aacaaggaaa tgtacctaaa attgcgtagc gagacactgt
     1441 tgccacttac acagtataac aggtatcttg ctctatataa caagtacaag tatttcagtg
     1501 gagccttaga tactaccagc tatcgtgaag cagcttgctg ccacttagca aaggctctaa
     1561 atgactttag caactcaggt gctgatgttc tctaccaacc accacagaca tcaatcactt
     1621 ctgctgttct gcagtaatac ccaactttct tgtacaaagt tggcattata agaaagcatt
     1681 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     1741 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     1801 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1861 gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
     1921 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     1981 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     2041 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     2101 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     2161 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     2221 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     2281 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     2341 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     2401 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     2461 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     2521 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     2581 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     2641 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     2701 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     2761 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     2821 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     2881 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     2941 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     3001 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     3061 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3241 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3361 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3421 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3481 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3541 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3661 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3721 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3781 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3841 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     3901 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     3961 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     4021 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     4081 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     4141 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     4201 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt
     4261 taacgctagc atggatgttt tcccagtcac gacgtt
//
