LOCUS       sequence-420611-        6611 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 6611)
  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 6611)
  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: pEntry_SARS-CoV-1-Spike
FEATURES             Location/Qualifiers
     source          1..6611
                     /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    3873..3884
                     /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(3954..4053)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4070..4089)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4071..4089)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4094..4110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4094..4110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4541..5332
                     /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      5425..6013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5914..5933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6167..6184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6343..6370)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6462..6548)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6597..6611,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6611,1..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 aaaaagcaga agggccgtca aggccaggat ccatgggatc ctttattttc ttattatttc
      181 ttactctcac tagtggtagt gaccttgacc ggtgcaccac ttttgatgat gttcaagctc
      241 ctaattacac tcaacatact tcatctatga ggggggttta ctatcctgat gaaattttta
      301 gatcagacac tctttattta actcaggatt tatttcttcc attttattct aatgttacag
      361 ggtttcatac tattaatcat acgtttggca accctgtcat accttttaag gatggtattt
      421 attttgctgc cacagagaaa tcaaatgttg tccgtggttg ggtttttggt tctaccatga
      481 acaacaagtc acagtcggtg attattatta acaattctac taatgttgtt atacgagcat
      541 gtaactttga attgtgtgac aaccctttct ttgctgtttc taaacccatg ggtacacaga
      601 cacatactat gatattcgat aatgcattta attgcacttt cgagtacata tctgatgcct
      661 tttcgcttga tgtttcagaa aagtcaggta attttaaaca cttacgagag tttgtgttta
      721 aaaataaaga tgggtttctc tatgtttata agggctatca acctatagat gtagttcgtg
      781 atctaccttc tggttttaac actttgaaac ctatttttaa gttgcctctt ggtattaaca
      841 ttacaaattt tagagccatt cttacagcct tttcacctgc tcaagacatt tggggcacgt
      901 cagctgcagc ctattttgtt ggctatttaa agccaactac atttatgctc aagtatgatg
      961 aaaatggtac aatcacagat gctgttgatt gttctcaaaa tccacttgct gaactcaaat
     1021 gctctgttaa gagctttgag attgacaaag gaatttacca gacctctaat ttcagggttg
     1081 ttccctcagg agatgttgtg agattcccta atattacaaa cttgtgtcct tttggagagg
     1141 tttttaatgc tactaaattc ccttctgtct atgcatggga gagaaaaaaa atttctaatt
     1201 gtgttgctga ttactctgtg ctctacaact caacattttt ttcaaccttt aagtgctatg
     1261 gcgtttctgc cactaagttg aatgatcttt gcttctccaa tgtctatgca gattcttttg
     1321 tagtcaaggg agatgatgta agacaaatag cgccaggaca aactggtgtt attgctgatt
     1381 ataattataa attgccagat gatttcatgg gttgtgtcct tgcttggaat actaggaaca
     1441 ttgatgctac ttcaactggt aattataatt ataaatatag gtatcttaga catggcaagc
     1501 ttaggccctt tgagagagac atatctaatg tgcctttctc ccctgatggc aaaccttgca
     1561 ccccacctgc tcttaattgt tattggccat taaatgatta tggtttttac accactactg
     1621 gcattggcta ccaaccttac agagttgtag tactttcttt tgaactttta aatgcaccgg
     1681 ccacggtttg tggaccaaaa ttatccactg accttattaa gaaccagtgt gtcaatttta
     1741 attttaatgg actcactggt actggtgtgt taactccttc ttcaaagaga tttcaaccat
     1801 ttcaacaatt tggccgtgat gtttctgatt tcactgattc cgttcgagat cctaaaacat
     1861 ctgaaatatt agacatttca ccttgcgctt ttgggggtgt aagtgtaatt acacctggaa
     1921 caaatgcttc atctgaagtt gctgttctat atcaagatgt taactgcact gatgtttcta
     1981 cagcaattca tgcagatcaa ctcacaccag cttggcgcat atattctact ggaaacaatg
     2041 tattccagac tcaagcaggc tgtcttatag gagctgagca tgtcgacact tcttatgagt
     2101 gcgacattcc tattggagct ggcatttgtg ctagttacca tacagtttct ttattacgta
     2161 gtactagcca aaaatctatt gtggcttata ctatgtcttt aggtgctgat agttcaattg
     2221 cttactctaa taacaccatt gctataccta ctaacttttc aattagcatt actacagaag
     2281 taatgcctgt ttctatggct aaaacctccg tagattgtaa tatgtacatc tgcggagatt
     2341 ctactgaatg tgctaatttg cttctccaat atggtagctt ttgcacacaa ctaaatcgtg
     2401 cactctcagg tattgctgct gaacaggatc gcaacacacg tgaagtgttc gctcaagtca
     2461 aacaaatgta caaaacccca actttgaaat attttggtgg ttttaatttt tcacaaatat
     2521 tacctgaccc tctaaagcca actaagaggt cttttattga ggacttgctc tttaataagg
     2581 tgacactcgc tgatgctggc ttcatgaagc aatatggcga atgcctaggt gatattaatg
     2641 ctagagatct catttgtgcg cagaagttca atggacttac agtgttgcca cctctgctca
     2701 ctgatgatat gattgctgcc tacactgctg ctctagttag tggtactgcc actgctggat
     2761 ggacatttgg tgctggcgct gctcttcaaa taccttttgc tatgcaaatg gcatataggt
     2821 tcaatggcat tggagttacc caaaatgttc tctatgagaa ccaaaaacaa atcgccaacc
     2881 aatttaacaa ggcgattagt caaattcaag aatcacttac aacaacatca actgcattgg
     2941 gcaagctgca agacgttgtt aaccagaatg ctcaagcatt aaacacactt gttaaacaac
     3001 ttagctctaa ttttggtgca atttcaagtg tgctaaatga tatcctttcg cgacttgata
     3061 aagtcgaggc ggaggtacaa attgacaggt taattacagg cagacttcaa agccttcaaa
     3121 cctatgtaac acaacaacta atcagggctg ctgaaatcag ggcttctgct aatcttgctg
     3181 ctactaaaat gtctgagtgt gttcttggac aatcaaaaag agttgacttt tgtggaaagg
     3241 gctaccacct tatgtccttc ccacaagcag ccccgcatgg tgttgtcttc ctacatgtca
     3301 cgtatgtgcc atcccaggag aggaacttca ccacagcgcc agcaatttgt catgaaggca
     3361 aagcatactt ccctcgtgaa ggtgtttttg tgtttaatgg cacttcttgg tttattacac
     3421 agaggaactt cttttctcca caaataatta ctacagacaa tacatttgtc tcaggaaatt
     3481 gtgatgtcgt tattggcatc attaacaaca cagtttatga tcctctgcaa cctgagcttg
     3541 actcattcaa agaagagctg gacaagtact tcaaaaatca tacatcacca gatgttgatc
     3601 ttggcgacat ttcaggcatt aacgcttctg tcgtcaacat tcaaaaagaa attgaccgcc
     3661 tcaatgaggt cgctaaaaat ttaaatgaat cactcattga ccttcaagaa ttgggaaaat
     3721 atgagcaata tattaaatgg ccttggtatg tttggctcgg cttcattgct ggactaattg
     3781 ccatcgtcat ggttacaatc ttgctttgtt gcatgactag ttgttgcagt tgcctcaagg
     3841 gtgcatgctc ttgtggttct tgctgcaagt ttgatgagga tgactctgag ccagttctca
     3901 agggtgtcaa attacattac acacttaatt aagacgcgat cgcggcctca tgggcccagc
     3961 tttcttgtac aaagttggca ttataaaaaa taattgctca tcaatttgtt gcaacgaaca
     4021 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     4081 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     4141 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     4201 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     4261 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     4321 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     4381 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     4441 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     4501 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     4561 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     4621 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     4681 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     4741 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     4801 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     4861 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     4921 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     4981 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     5041 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     5101 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     5161 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     5221 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     5281 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     5341 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     5401 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5461 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5521 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5581 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5641 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5701 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5761 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5821 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5881 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5941 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6001 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6061 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6121 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6181 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6241 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6301 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     6361 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     6421 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     6481 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     6541 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     6601 gtcacgacgt t
//
