LOCUS       sequence-330957-        4111 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for inducible mammalian expression 
            using the TRE3G promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  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 4111)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 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-E31: attB4-TRE3Gp>-attB1r
FEATURES             Location/Qualifiers
     source          1..4111
                     /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"
     promoter        205..1416
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1253..1272
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    1418..1541
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1613..1632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1614..1632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1637..1653)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1637..1653)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2078..2869
                     /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      2962..3550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3451..3470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3704..3721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3880..3907)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3999..4085)
                     /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 agttggcctc cgcgccgggt tttggcgcct cccgcgggcg
      241 cccccctcct cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc
      301 cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc
      361 agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct
      421 ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat
      481 ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag
      541 ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact
      601 tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg
      661 gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc
      721 ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga
      781 agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc
      841 aagaacccaa ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg
      901 gctggggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
      961 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     1021 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     1081 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     1141 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     1201 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     1261 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     1321 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc
     1381 gctaaattct ggccgttttt ggcttttttg ttagacgcaa ctttgtacaa aaaagttgaa
     1441 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
     1501 actacataat actgtaaaac acaacatatg cagtcactat gaaccaacta cttagatggt
     1561 attagtgacc tgtagaattc gagctctaga gctgcagggc ggccgcgata tcccctatag
     1621 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     1681 ctgatgttac attgcacaag atcggtaccc ggggatcctc tagaccagcc aggacagaaa
     1741 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     1801 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     1861 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     1921 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     1981 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     2041 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     2101 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     2161 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     2221 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     2281 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     2341 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     2401 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     2461 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     2521 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     2581 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     2641 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     2701 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     2761 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     2821 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     2881 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3601 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3661 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3721 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3781 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     3841 tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
     3901 gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     3961 ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     4021 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt
     4081 atttgatgcc tggcagttcc ctactctcgc g
//
