LOCUS       sequence-333051-        6555 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 6555)
  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 6555)
  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_nostop
FEATURES             Location/Qualifiers
     source          1..6555
                     /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(3898..3997)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4014..4033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4015..4033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4038..4054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4038..4054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4485..5276
                     /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      5369..5957
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5858..5877
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6111..6128
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6287..6314)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6406..6492)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6541..6555,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6555,1..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 tacacatacc
     3901 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     3961 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     4021 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     4081 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     4141 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     4201 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     4261 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     4321 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     4381 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     4441 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     4501 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     4561 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     4621 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     4681 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     4741 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     4801 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     4861 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     4921 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     4981 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     5041 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     5101 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     5161 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     5221 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     5281 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     5341 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5401 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5461 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5521 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5581 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5641 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5701 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5761 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5821 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5881 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5941 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6001 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6061 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6121 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6181 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6241 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     6301 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     6361 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     6421 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     6481 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     6541 cccagtcacg acgtt
//
