LOCUS       sequence-333053-        6558 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 6558)
  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 6558)
  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 S_stop
FEATURES             Location/Qualifiers
     source          1..6558
                     /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    3846..3857
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(3901..4000)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4017..4036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4018..4036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4041..4057)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4041..4057)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4488..5279
                     /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      5372..5960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5861..5880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6114..6131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6290..6317)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6409..6495)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6544..6558,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6558,1..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 catgtttatt ttcttattat ttcttactct cactagtggt agtgaccttg
      181 accggtgcac cacttttgat gatgttcaag ctcctaatta cactcaacat acttcatcta
      241 tgaggggggt ttactatcct gatgaaattt ttagatcaga cactctttat ttaactcagg
      301 atttatttct tccattttat tctaatgtta cagggtttca tactattaat catacgtttg
      361 gcaaccctgt catacctttt aaggatggta tttattttgc tgccacagag aaatcaaatg
      421 ttgtccgtgg ttgggttttt ggttctacca tgaacaacaa gtcacagtcg gtgattatta
      481 ttaacaattc tactaatgtt gttatacgag catgtaactt tgaattgtgt gacaaccctt
      541 tctttgctgt ttctaaaccc atgggtacac agacacatac tatgatattc gataatgcat
      601 ttaattgcac tttcgagtac atatctgatg ccttttcgct tgatgtttca gaaaagtcag
      661 gtaattttaa acacttacga gagtttgtgt ttaaaaataa agatgggttt ctctatgttt
      721 ataagggcta tcaacctata gatgtagttc gtgatctacc ttctggtttt aacactttga
      781 aacctatttt taagttgcct cttggtatta acattacaaa ttttagagcc attcttacag
      841 ccttttcacc tgctcaagac atttggggca cgtcagctgc agcctatttt gttggctatt
      901 taaagccaac tacatttatg ctcaagtatg atgaaaatgg tacaatcaca gatgctgttg
      961 attgttctca aaatccactt gctgaactca aatgctctgt taagagcttt gagattgaca
     1021 aaggaattta ccagacctct aatttcaggg ttgttccctc aggagatgtt gtgagattcc
     1081 ctaatattac aaacttgtgt ccttttggag aggtttttaa tgctactaaa ttcccttctg
     1141 tctatgcatg ggagagaaaa aaaatttcta attgtgttgc tgattactct gtgctctaca
     1201 actcaacatt tttttcaacc tttaagtgct atggcgtttc tgccactaag ttgaatgatc
     1261 tttgcttctc caatgtctat gcagattctt ttgtagtcaa gggagatgat gtaagacaaa
     1321 tagcgccagg acaaactggt gttattgctg attataatta taaattgcca gatgatttca
     1381 tgggttgtgt ccttgcttgg aatactagga acattgatgc tacttcaact ggtaattata
     1441 attataaata taggtatctt agacatggca agcttaggcc ctttgagaga gacatatcta
     1501 atgtgccttt ctcccctgat ggcaaacctt gcaccccacc tgctcttaat tgttattggc
     1561 cattaaatga ttatggtttt tacaccacta ctggcattgg ctaccaacct tacagagttg
     1621 tagtactttc ttttgaactt ttaaatgcac cggccacggt ttgtggacca aaattatcca
     1681 ctgaccttat taagaaccag tgtgtcaatt ttaattttaa tggactcact ggtactggtg
     1741 tgttaactcc ttcttcaaag agatttcaac catttcaaca atttggccgt gatgtttctg
     1801 atttcactga ttccgttcga gatcctaaaa catctgaaat attagacatt tcaccttgcg
     1861 cttttggggg tgtaagtgta attacacctg gaacaaatgc ttcatctgaa gttgctgttc
     1921 tatatcaaga tgttaactgc actgatgttt ctacagcaat tcatgcagat caactcacac
     1981 cagcttggcg catatattct actggaaaca atgtattcca gactcaagca ggctgtctta
     2041 taggagctga gcatgtcgac acttcttatg agtgcgacat tcctattgga gctggcattt
     2101 gtgctagtta ccatacagtt tctttattac gtagtactag ccaaaaatct attgtggctt
     2161 atactatgtc tttaggtgct gatagttcaa ttgcttactc taataacacc attgctatac
     2221 ctactaactt ttcaattagc attactacag aagtaatgcc tgtttctatg gctaaaacct
     2281 ccgtagattg taatatgtac atctgcggag attctactga atgtgctaat ttgcttctcc
     2341 aatatggtag cttttgcaca caactaaatc gtgcactctc aggtattgct gctgaacagg
     2401 atcgcaacac acgtgaagtg ttcgctcaag tcaaacaaat gtacaaaacc ccaactttga
     2461 aatattttgg tggttttaat ttttcacaaa tattacctga ccctctaaag ccaactaaga
     2521 ggtcttttat tgaggacttg ctctttaata aggtgacact cgctgatgct ggcttcatga
     2581 agcaatatgg cgaatgccta ggtgatatta atgctagaga tctcatttgt gcgcagaagt
     2641 tcaatggact tacagtgttg ccacctctgc tcactgatga tatgattgct gcctacactg
     2701 ctgctctagt tagtggtact gccactgctg gatggacatt tggtgctggc gctgctcttc
     2761 aaataccttt tgctatgcaa atggcatata ggttcaatgg cattggagtt acccaaaatg
     2821 ttctctatga gaaccaaaaa caaatcgcca accaatttaa caaggcgatt agtcaaattc
     2881 aagaatcact tacaacaaca tcaactgcat tgggcaagct gcaagacgtt gttaaccaga
     2941 atgctcaagc attaaacaca cttgttaaac aacttagctc taattttggt gcaatttcaa
     3001 gtgtgctaaa tgatatcctt tcgcgacttg ataaagtcga ggcggaggta caaattgaca
     3061 ggttaattac aggcagactt caaagccttc aaacctatgt aacacaacaa ctaatcaggg
     3121 ctgctgaaat cagggcttct gctaatcttg ctgctactaa aatgtctgag tgtgttcttg
     3181 gacaatcaaa aagagttgac ttttgtggaa agggctacca ccttatgtcc ttcccacaag
     3241 cagccccgca tggtgttgtc ttcctacatg tcacgtatgt gccatcccag gagaggaact
     3301 tcaccacagc gccagcaatt tgtcatgaag gcaaagcata cttccctcgt gaaggtgttt
     3361 ttgtgtttaa tggcacttct tggtttatta cacagaggaa cttcttttct ccacaaataa
     3421 ttactacaga caatacattt gtctcaggaa attgtgatgt cgttattggc atcattaaca
     3481 acacagttta tgatcctctg caacctgagc ttgactcatt caaagaagag ctggacaagt
     3541 acttcaaaaa tcatacatca ccagatgttg atcttggcga catttcaggc attaacgctt
     3601 ctgtcgtcaa cattcaaaaa gaaattgacc gcctcaatga ggtcgctaaa aatttaaatg
     3661 aatcactcat tgaccttcaa gaattgggaa aatatgagca atatattaaa tggccttggt
     3721 atgtttggct cggcttcatt gctggactaa ttgccatcgt catggttaca atcttgcttt
     3781 gttgcatgac tagttgttgc agttgcctca agggtgcatg ctcttgtggt tcttgctgca
     3841 agtttgatga ggatgactct gagccagttc tcaagggtgt caaattacat tacacataat
     3901 acccaacttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     3961 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     4021 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     4081 atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc
     4141 aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga
     4201 aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca
     4261 agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg
     4321 gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc
     4381 gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca
     4441 acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg
     4501 tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg
     4561 aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac
     4621 acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag
     4681 ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg
     4741 ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc
     4801 gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag
     4861 ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct
     4921 tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg
     4981 cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa
     5041 atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga
     5101 aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac
     5161 ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc
     5221 agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac
     5281 cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt
     5341 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5401 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5461 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5521 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5581 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5641 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5701 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5761 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5821 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     5881 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     5941 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6001 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6061 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     6121 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     6181 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     6241 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     6301 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     6361 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     6421 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     6481 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
     6541 tttcccagtc acgacgtt
//
