LOCUS       sequence-330930-        2996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            Myc-tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2996)
  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 2996)
  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: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C514-E26: attB5r-RU5u-myc-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..2996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(44..167)
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             171..439
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             452..481
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    482..606
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(678..697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(679..697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(702..718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(702..718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1143..1934
                     /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      2027..2615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2516..2535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2750..2777)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2869..2955)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agctcgaatt ctacagtcac taataccatc taagtagttg attcatagtg actgcatatg
       61 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
      121 atttatatca ttttacgttt ctcgttcaac ttttgtatac aaagtggcga ggggctcgca
      181 tctctccttc acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg
      241 ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa
      301 agctcaggtc gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc
      361 ggctctccac gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg
      421 ttctgcgccg ttacagatca ccatggctag cgaacaaaag ctcatatctg aagaggacct
      481 aacaactttg tacaaaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt
      541 aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac atatgcagtc
      601 actatgaacc aactacttag atggtattag tgacctgtag aattcgagct ctagagctgc
      661 agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
      721 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagatcgg tacccgggga
      781 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
      841 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
      901 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
      961 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1021 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     1081 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     1141 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1201 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1261 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     1321 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     1381 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     1441 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     1501 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     1561 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     1621 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     1681 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     1741 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     1801 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     1861 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     1921 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     1981 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2041 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2101 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2161 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2221 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2281 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2341 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2401 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2461 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2521 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2581 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2641 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2701 ctgtggataa ccgtattacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
     2761 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
     2821 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
     2881 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
     2941 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta ccatgg
//
