LOCUS       sequence-330926-        3538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            mClover3 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3538)
  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 3538)
  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: C232-E29: attB2r-mClover3-attB3
FEATURES             Location/Qualifiers
     source          1..3538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(152..275)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(999..1094)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1505..2296
                     /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      2389..2977
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2878..2897
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3131..3148
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3307..3334)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3426..3512)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgatat ccttaagctc gggccctgca gctctagagc tcgaattcta
      121 caggtcacta ataccatcta agtagttggt tcatagtgac tgcatatgtt gtgttttaca
      181 gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat ttatatcatt
      241 ttacgtttct cgttcaactt tcttgtacaa agtgggcggt gtgtcgaagg gcgaagaact
      301 ctttaccgga gtggtgccga ttctggtgga actggacgga gatgtcaacg gtcataagtt
      361 ctccgtccgg ggagaaggag agggcgatgc cactaacggg aagctcacgc tgaaattcat
      421 ctgcaccact gggaagcttc cggtgccctg gcctaccctc gtcaccacct tcggttacgg
      481 ggtggcctgt ttctcacggt acccagacca catgaaacaa catgacttct ttaagtccgc
      541 aatgcccgag ggatacgtgc aggaacgcac catcagcttc aaggacgacg gcacttacaa
      601 gactagagcc gaagtcaagt tcgagggaga tacactcgtg aacaggatcg aactgaaggg
      661 cattgatttc aaggaggacg gcaacattct gggccacaag ttggagtaca acttcaattc
      721 ccactacgtg tacatcaccg cggacaagca gaagaactgc atcaaggcca acttcaagat
      781 ccgccacaac gtggaggatg gatcggtcca gctggcggat cactaccagc aaaacacccc
      841 cattggcgac ggtcctgtgc tgctgcctga caaccactac ctgagccacc agtccaagtt
      901 gtccaaagac ccgaatgaga agcgggacca catggtgctg ctggaattcg tgactgccgc
      961 tggaatcacc catgggatgg acgagctgta taagtaatca actttattat acaaagttgg
     1021 cattataaaa aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1081 taaaatcatt atttggagct ccatggtagc gttaacgcgg ccgcgatatc ggtacccggg
     1141 gatcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     1201 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     1261 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     1321 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     1381 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     1441 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     1501 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     1561 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     1621 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     1681 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     1741 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     1801 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     1861 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     1921 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     1981 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     2041 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     2101 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     2161 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     2221 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     2281 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     2341 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     2401 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     2461 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     2521 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     2581 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     2641 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     2701 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     2761 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     2821 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     2881 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     2941 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3001 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     3061 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     3121 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     3181 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     3241 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     3301 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     3361 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     3421 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     3481 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcg
//
