LOCUS       sequence-330613-        2402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4-attB5 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 2402)
  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 2402)
  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: C455-E01: attB4-stuffer-attB5
FEATURES             Location/Qualifiers
     source          1..2402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..102
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     195..214
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(256..351)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      399..485
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      577..604
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(739..1327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(819..838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1470..2279)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2193..2212
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2271..2290
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
       61 agcaatgctt ttttataatg ccaactttgt atacaaaagt tgtccccagg atgttgccgt
      121 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
      181 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
      241 cgtagccttc ggggccaact tttctataca aagttggcat tataaaaaag cattgcttat
      301 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca
      361 tggtagcgtt aacgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
      421 cggaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
      481 caaatccgcc gggagcggat ttgaacgttg tgaagcaacg gcccggaggg tggcgggcag
      541 gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa ggccatcctg acggatggcc
      601 tttttgcgtt tctacaaact cttcctggct agcggtaata cggttatcca cagaatcagg
      661 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      721 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      781 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      841 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      901 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      961 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1021 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1081 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1141 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1201 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1261 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1321 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1381 acgttaaggg attttggtca tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc
     1441 cggtgttaca accaattaac caattctgat tagaaaaact catcgagcat caaatgaaac
     1501 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     1561 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     1621 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     1681 tcaagtgaga aatcaccatg agtgacgact gaatccggcg agaatggcaa aagtttatgc
     1741 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     1801 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     1861 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     1921 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgtttttccg
     1981 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     2041 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     2101 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag
     2161 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     2221 tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata
     2281 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt
     2341 ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccctgcgcc ctgcagctct
     2401 ag
//
