LOCUS       sequence-330548-        3618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r/attB1r entry clone for generation of N-terminal 
            iRFP670 fusion proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3618)
  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 3618)
  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: C512-E34: attB5r-iRFP670-nostop-attB1r
FEATURES             Location/Qualifiers
     source          1..3618
                     /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"
     protein_bind    1245..1369
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1441..1460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1442..1460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1465..1481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1465..1481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1906..2697
                     /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      2790..3378
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3279..3298
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3513..3540)
                     /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 caaagtggac catggccagg aaggtcgacc tgaccagctg cgatagagag cccatccata
      361 tcccgggatc cattcagccg tgcggatgcc tcctggcctg tgacgctcag gcagtgcgga
      421 tcacccgcat tactgagaat gccggagcct tcttcggacg cgagactccc agagtgggcg
      481 aactccttgc cgactacttt ggcgaaacgg aagcccatgc gctgcggaac gcgttggcgc
      541 agtcgtccga cccaaagagg cctgccctga tcttcggatg gcgcgacggt cttaccgggc
      601 gcaccttcga tatttccctg caccggcacg acggaacctc aatcatcgag ttcgaaccag
      661 ctgctgcgga acaggccgat aacccgctgc ggctgactcg gcaaatcatt gcccgcacca
      721 aggagcttaa atccctggag gagatggcgg ccagagtccc tagatacctc caggccatgc
      781 tcgggtatca ccgcgtgatg ctgtaccggt tcgcggacga cggttcaggg atggtcattg
      841 gagaagccaa gaggagcgac ctggaatcct tcctgggtca gcactttccc gcctcgttgg
      901 tgccgcaaca agccaggctc ctgtacctga agaacgccat ccgagtggtg tcagacagcc
      961 gcggcatcag ctcccggatc gtgcctgaac atgatgccag cggagctgcc ttggatctct
     1021 ctttcgccca cctgagatcc atctccccct gtcacctgga gtttctgcgg aacatgggcg
     1081 tgtcggcatc catgtcgctg tccatcatca ttgacggcac actctggggc ctgattatct
     1141 gccaccacta cgaacctcgg gcagtgccga tggcccagag agtggccgct gagatgttcg
     1201 cagacttcct gtccctgcac ttcaccgcgg cccatcacca gcgcacaact ttgtacaaaa
     1261 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     1321 aaaaacagac tacataatac tgtaaaacac aacatatgca gtcactatga accaactact
     1381 tagatggtat tagtgacctg tagaattcga gctctagagc tgcagggcgg ccgcgatatc
     1441 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     1501 caaaatctct gatgttacat tgcacaagat cggtacccgg ggatcctcta gaccagccag
     1561 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
     1621 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     1681 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     1741 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     1801 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     1861 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
     1921 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     1981 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     2041 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     2101 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     2161 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     2221 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     2281 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     2341 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     2401 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     2461 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     2521 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     2581 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     2641 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     2701 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     2761 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2821 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2881 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2941 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3001 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3061 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3121 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3181 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3241 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3301 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3361 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3421 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3481 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3541 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3601 gcttcacaac gttcaaat
//
