LOCUS       sequence-333712-        3255 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 3255)
  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 3255)
  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 ORF3b_stop
FEATURES             Location/Qualifiers
     source          1..3255
                     /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(598..697)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(714..733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(715..733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(738..754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(738..754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1185..1976
                     /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      2069..2657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2558..2577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2811..2828
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2987..3014)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3106..3192)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3241..3255,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3255,1..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 catgatgcca actactttgt ttgctggcac acacataact atgactactg
      181 tataccatat aacagtgtca cagatacaat tgtcgttact gaaggtgacg gcatttcaac
      241 accaaaactc aaagaagact accaaattgg tggttattct gaggataggc actcaggtgt
      301 taaagactat gtcgttgtac atggctattt caccgaagtt tactaccagc ttgagtctac
      361 acaaattact acagacactg gtattgaaaa tgctacattc ttcatcttta acaagcttgt
      421 taaagaccca ccgaatgtgc aaatacacac aatcgacggc tcttcaggag ttgctaatcc
      481 agcaatggat acaatttatg atgagccgac gacgactact agcgtgcctt tgtaagcaca
      541 agaaagtgag tacgaactta tgtactcatt cgtttcggaa gaaacaggta cgttaatacc
      601 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
      661 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
      721 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
      781 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
      841 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
      901 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
      961 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     1021 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     1081 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     1141 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     1201 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     1261 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     1321 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     1381 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     1441 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     1501 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     1561 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     1621 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     1681 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     1741 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     1801 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     1861 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     1921 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     1981 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     2041 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2101 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2161 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2221 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2281 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2341 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2401 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2461 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2521 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2581 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2641 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2701 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2761 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2821 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2881 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2941 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     3001 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     3061 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     3121 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     3181 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     3241 cccagtcacg acgtt
//
