LOCUS       sequence-333332-        3004 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 3004)
  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 3004)
  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-E07: attB2r-SV40polyA-attB3
FEATURES             Location/Qualifiers
     source          1..3004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(109..232)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    363..497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(400..419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     454..473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(499..594)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             964..1755
                     /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      1847..2435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2336..2355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2589..2606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2765..2792)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2884..2970)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta aaattcgagc
       61 tcggtccgag gatcctacag gtcactaata ccatctaagt agttgattca tagtgactgg
      121 atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata
      181 ttgatattta tatcacctta cgtttctcgt tcagctttct tgtacaaagt gggcagcttg
      241 tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
      301 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      361 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      421 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      481 cttatcatgt ctggatctca actttattat acaaagttgg cattatagga aagcattgct
      541 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgcatgc
      601 aagcttagat cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
      661 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
      721 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
      781 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactggg
      841 gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg
      901 atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac
      961 attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1021 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1081 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1141 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     1201 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     1261 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     1321 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     1381 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     1441 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     1501 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     1561 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     1621 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     1681 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1741 gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg
     1801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag atcttcttga gatccttttt
     1861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1921 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1981 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2041 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2101 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2161 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2221 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2281 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2341 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2401 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2461 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2521 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2581 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2641 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2701 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     2761 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     2821 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     2881 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     2941 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     3001 cgct
//
