LOCUS       sequence-333750-        4554 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 4554)
  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 4554)
  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 HCoV-HKU1 NSP2_nostop
FEATURES             Location/Qualifiers
     source          1..4554
                     /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(1897..1996)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2013..2032)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2014..2032)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2037..2053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2037..2053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2484..3275
                     /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      3368..3956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3857..3876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4110..4127
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4286..4313)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4405..4491)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4540..4554,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4554,1..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 catgattaaa cccgtacttt ttgtagacca gtatggttgt gactattctg
      181 gtaaattagc agattgtctt caagcttatg gtcattattc tttgcaagat atgagacaaa
      241 agcagtctgt atggcttgcc aattgtgact ttgatattgt agtggcttgg catgtagttc
      301 gtgattcacg atttgttatg cgcctgcaga ctatagctac tatttgtggt attaaatatg
      361 ttgcacaacc tacagaagat gtagtagatg gagatgtagt tatacgtgaa cctgtacatt
      421 tattatctgc tgatgcaata gttttaaagc ttcctagttt gatgaaagtt atgactcata
      481 tggatgattt ttctattaaa tctatatata atgttgattt gtgtgattgt ggttttgtta
      541 tgcagtatgg ttatgtagat tgttttaatg ataattgtga tttttatggt tgggtttcag
      601 gtaatatgat ggatggtttt tcttgtccat tgtgttgtac agtttatgac tctagcgaag
      661 ttaaagccca atcatctggt gttattcctg aaaatcctgt gttatttact aatagtactg
      721 atactgttaa ccatgattct tttaatttgt atggttattc tgtcacacca tttggttctt
      781 gtatatattg gtcgccgcgt cctggattgt ggattcctat aattaaatct tcagtcaagt
      841 cttatgatga tttggtttat tcaggtgtag taggttgtaa atctattgtt aaagaaactg
      901 ctcttattac tcatgcactt tacttagatt atgttcaatg taagtgtggt aatcttgaac
      961 aaaatcatat tcttggcgtt aataattctt ggtgtaggca actgttgctt aatagaggtg
     1021 attataatat gcttctaaaa aatattgact tgtttgttaa gcgtcgtgct gattttgctt
     1081 gcaagtttgc agtttgtgga gatggttttg tacctttttt actagatggt ttaattcccc
     1141 gtagttatta tctaattcag agtggtattt tctttacatc tttgatgtct caattttcac
     1201 aagaagtttc tgatatgtgt ttaaaaatgt gtattttgtt tatggacaga gtttcagttg
     1261 ctacatttta tatagagcat tatgttaata ggttggttac tcaatttaag ttattgggta
     1321 ctacacttgt taataaaatg gttaattggt ttaataccat gttagatgct agtgcacctg
     1381 ctacaggctg gcttctttac caattattga atggtctttt tgtagtatct caagccaact
     1441 ttaattttgt tgctttaata cctgattatg ctaaaatttt agttaataaa ttttacactt
     1501 tttttaagtt attattagag tgtgttacag ttgatgtttt aaaagatatg cctgttctta
     1561 aaactattaa tggtttagtt tgtattgtag gcaataagtt ttataacgtt agtacagggt
     1621 taattcctgg ttttgtttta ccatgtaatg cacaggaaca acaaatttat ttttttgaag
     1681 gcgttgcaga atctgttata gtagaagatg atgttattga gaatgtcaaa tcttctttat
     1741 catcttatga gtattgtcaa ccacctaaat ctgtagaaaa aatttgtatt atagataata
     1801 tgtacatggg taagtgtggt gataaatttt tccctattgt catgaatgat aaaaatattt
     1861 gtcttttaga tcaggcttgg cgttttccat gtgcataccc aactttcttg tacaaagttg
     1921 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1981 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     2041 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2101 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     2161 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     2221 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     2281 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     2341 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     2401 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     2461 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     2521 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     2581 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     2641 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     2701 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     2761 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     2821 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     2881 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     2941 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     3001 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     3061 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     3121 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     3181 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     3241 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     3301 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3481 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4021 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4081 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4141 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4201 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     4261 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     4321 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     4381 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4441 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     4501 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
