LOCUS       sequence-330602-        3002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            V5-tagged fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3002)
  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 3002)
  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-E24: attB5r-RU5u-V5-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3002
                     /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             449..490
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    488..612
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(684..703)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(685..703)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(708..724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(708..724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1149..1940
                     /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      2033..2621
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2522..2541
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2756..2783)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2875..2961)
                     /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 ttacagatcg gcaccatggg taagcctatc cctaaccctc tcctcggtct
      481 cgattctaca actttgtaca aaaaagttga acgagaaacg taaaatgata taaatatcaa
      541 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
      601 gcagtcacta tgaaccaact acttagatgg tattagtgac ctgtagaatt cgagctctag
      661 agctgcaggg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
      721 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gatcggtacc
      781 cggggatcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg
      841 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
      901 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
      961 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     1021 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     1081 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     1141 taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     1201 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     1261 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     1321 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     1381 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     1441 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     1501 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     1561 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     1621 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     1681 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     1741 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     1801 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     1861 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     1921 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     1981 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2041 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2101 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2161 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2221 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2281 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2341 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2401 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2461 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2521 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2581 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2641 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2701 ctgattctgt ggataaccgt attaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     2761 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     2821 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     2881 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     2941 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctaccat
     3001 gg
//
