LOCUS       sequence-330995-        2990 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            FLAG-tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2990)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 2990)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C514-E09: attB5r-RU5u-FLAG-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..2990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(14..100)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    complement(185..308)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             312..580
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             590..613
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    617..741
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(813..832)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(814..832)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(837..853)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(837..853)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1278..2069
                     /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      2162..2750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2651..2670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2885..2912)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
       61 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
      121 ctcgcgttaa cgctaccatg gagctcgaat tctacagtca ctaataccat ctaagtagtt
      181 gattcatagt gactgcatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
      241 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa cttttgtata
      301 caaagtggcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
      361 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
      421 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
      481 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
      541 acgtctttgt ttcgttttct gttctgcgcc gttacagatc accatgggcg attataagga
      601 tgacgatgac aagggcacaa ctttgtacaa aaaagttgaa cgagaaacgt aaaatgatat
      661 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
      721 acaacatatg cagtcactat gaaccaacta cttagatggt attagtgacc tgtagaattc
      781 gagctctaga gctgcagggc ggccgcgata tcccctatag tgagtcgtat tacatggtca
      841 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
      901 atcggtaccc ggggatcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac
      961 ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat
     1021 aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca
     1081 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     1141 actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg
     1201 tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta
     1261 aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag
     1321 aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg
     1381 gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga
     1441 ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg
     1501 cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg
     1561 acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca
     1621 atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc
     1681 tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg
     1741 atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact
     1801 cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt
     1861 acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc
     1921 gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag
     1981 aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag
     2041 gcgagatcac caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta
     2101 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2161 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2221 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2701 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2761 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2821 cgttatcccc tgattctgtg gataaccgta ttaccgctag ccaggaagag tttgtagaaa
     2881 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     2941 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
//
