LOCUS       sequence-330537-        3239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for mammalian expression using the 
            SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3239)
  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 3239)
  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: C413-E26: attB4-SV40p>-attB1r
FEATURES             Location/Qualifiers
     source          1..3239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    110..205
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(212..232)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        320..530
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        342..538
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      389..524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    546..669
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(741..760)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(742..760)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(765..781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(765..781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1206..1997
                     /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      2090..2678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2579..2598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2832..2849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3008..3035)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3127..3213)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgatcg ggcccgcgtt aacgctacca tggagctcca aataatgatt
      121 ttattttgac tgatagtgac ctgttcgttg caacaaattg ataagcaatg cttttttata
      181 atgccaactt tgtatagaaa agttggcctg tggaatgtgt gtcagttagg gtgtggaaag
      241 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaagg
      301 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      361 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      421 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      481 ctctgagcta ttccagaagt agtgaagagg cttttttgga ggcctaggct tttgcaaaaa
      541 gctggcaact ttgtacaaaa aagttgaacg agaaacgtaa aatgatataa atatcaatat
      601 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatgca
      661 gtcactatga accaactact tagatggtat tagtgacctg tagaattcga gctctagagc
      721 tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
      781 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat cggtacccgg
      841 ggatcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
      901 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
      961 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     1021 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     1081 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     1141 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     1201 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     1261 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     1321 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     1381 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     1441 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     1501 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     1561 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     1621 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     1681 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     1741 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     1801 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     1861 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     1921 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     1981 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     2041 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2221 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2701 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2761 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2821 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     2881 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     2941 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     3001 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     3061 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     3121 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     3181 ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcg
//
