LOCUS       sequence-330929-        3526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone for construction of C-terminal 
            mCherry fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3526)
  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 3526)
  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-E06: attB2r-mCherry-attB3
FEATURES             Location/Qualifiers
     source          1..3526
                     /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"
     CDS             278..985
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPYDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(420..438)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     688..707
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(987..1082)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1493..2284
                     /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      2377..2965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2866..2885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3119..3136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3295..3322)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3414..3500)
                     /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 agtgggcgtg agcaagggcg aggaggataa
      301 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
      361 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      421 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
      481 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
      541 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
      601 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
      661 gcgcggcacc aacttcccct acgacggccc cgtaatgcag aagaagacca tgggctggga
      721 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
      781 gctgaagctg aaggacggag gccactacga cgctgaggtc aagaccacct acaaggccaa
      841 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
      901 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
      961 cggcatggac gagctgtaca agtaatcaac tttattatac aaagttggca ttataaaaaa
     1021 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     1081 ttggagctcc atggtagcgt taacgcggcc gcgatatcgg tacccgggga tcctctagac
     1141 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     1201 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     1261 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     1321 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     1381 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     1441 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     1501 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1561 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1621 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1681 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     1741 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     1801 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     1861 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     1921 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     1981 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     2041 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     2101 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     2161 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     2221 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     2281 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3061 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3121 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3181 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3241 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     3301 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     3361 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     3421 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     3481 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcg
//
