LOCUS       sequence-333331-        3036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB2r/attB3 entry clone containing a polyA sequence for 
            transcription termination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3036)
  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 3036)
  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: C238-E02: attB2r-BGHpolyA-attB3
FEATURES             Location/Qualifiers
     source          1..3036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(323..446)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(450..467)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    456..663
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(668..763)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1174..1965
                     /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      2058..2646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2547..2566
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2800..2817
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2976..3003)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa aattcgagct
      241 cggtccgata tccttaagct cgggccctgc agctctagag ctcgaattct acaggtcact
      301 aataccatct aagtagttgg ttcatagtga ctgcatatgt tgtgttttac agtattatgt
      361 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
      421 tcgttcaact ttcttgtaca aagtgggcgc ctcgactgtg ccttctagtt gccagccatc
      481 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
      541 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
      601 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
      661 ggataatcaa ctttattata caaagttggc attataaaaa agcattgctt atcaatttgt
      721 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggagctc catggtagcg
      781 ttaacgcggc cgcgatatcg gtacccgggg atcctctaga ccagccagga cagaaatgcc
      841 tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
      901 cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc
      961 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     1021 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     1081 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     1141 cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta
     1201 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     1261 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     1321 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     1381 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     1441 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     1501 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     1561 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     1621 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     1681 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     1741 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     1801 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     1861 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     1921 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca
     1981 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2041 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2101 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2161 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2221 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2281 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2341 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2401 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2461 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2521 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2581 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2641 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2701 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2761 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2821 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     2881 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
     2941 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3001 tctgcttagt ttgatgcctg gcagtttatg gcgggc
//
