LOCUS       sequence-330947-        2890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB5r-attB1r entry clone containing a 
            non-transcribed/translated region for use in generating negative 
            control vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2890)
  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 2890)
  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: C515-E01: attB5r-stuffer-attB1r
FEATURES             Location/Qualifiers
     source          1..2890
                     /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"
     primer_bind     complement(382..401)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    517..641
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(713..732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(714..732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(737..753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(737..753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1178..1969
                     /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      2062..2650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2551..2570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2785..2812)
                     /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 caaagtggta actgaataga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      361 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      421 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
      481 tatccggcct ttattcacat gcttgcccgc ctgtaaacaa ctttgtacaa aaaagttgaa
      541 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
      601 actacataat actgtaaaac acaacatatg cagtcactat gaaccaacta cttagatggt
      661 attagtgacc tgtagaattc gagctctaga gctgcagggc ggccgcgata tcccctatag
      721 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
      781 ctgatgttac attgcacaag atcggtaccc ggggatcctc tagaccagcc aggacagaaa
      841 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
      901 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
      961 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     1021 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     1081 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     1141 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     1201 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     1261 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     1321 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     1381 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     1441 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     1501 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     1561 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     1621 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     1681 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     1741 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     1801 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     1861 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     1921 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     1981 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2041 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2101 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2161 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2221 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2281 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2341 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2401 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2461 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2521 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2581 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2641 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2701 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag
     2761 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     2821 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     2881 acgttcaaat
//
