LOCUS       sequence-296469-        2961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway-compatible Entry vector, with insert of ORF3B gene's CDS 
            from SARS-CoV-2 isolate Wuhan-Hu-1. Does not contain stop codon. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2961)
  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 2961)
  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-CoV-2 ORF3B_nostop
FEATURES             Location/Qualifiers
     source          1..2961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(721..820)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(837..856)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(838..856)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(861..877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(861..877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1308..2099
                     /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      2192..2780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2681..2700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2934..2951
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcat ggcctactgc tggcgctgca ccagctgctg cttctccgag aggttccaga
      601 accacaaccc ccagaaggaa atggctacct ctaccctgca gggttgttct ctgtgcctgc
      661 agctggctgt ggtggtgtgc aacagcctgc tgaccccctt cgctagatgc tgctggcctt
      721 acccaacttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
      781 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
      841 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
      901 atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc
      961 aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga
     1021 aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca
     1081 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
     1141 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc
     1201 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     1261 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg
     1321 tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg
     1381 aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac
     1441 acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag
     1501 ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg
     1561 ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc
     1621 gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag
     1681 ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct
     1741 tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg
     1801 cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa
     1861 atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga
     1921 aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac
     1981 ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc
     2041 agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac
     2101 cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     2161 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2221 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2281 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2341 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2401 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2461 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2521 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2581 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2641 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2701 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2761 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2821 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2881 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2941 cagtgagcga ggaagcggaa g
//
