LOCUS       sequence-330533-        3408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB1r entry clone for mammalian expression using the 
            human PGK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3408)
  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 3408)
  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-E21: attB4-PGKp>-attB1r
FEATURES             Location/Qualifiers
     source          1..3408
                     /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        211..710
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(246..269)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     643..662
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    714..838
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(910..929)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(911..929)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(934..950)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(934..950)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1375..2166
                     /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      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2748..2767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3001..3018
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3177..3204)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3296..3382)
                     /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 agttggcacc gggtagggga ggcgcttttc ccaaggcagt
      241 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
      301 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
      361 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
      421 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
      481 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
      541 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
      601 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
      661 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg accacaactt
      721 tgtacaaaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
      781 ttttgcataa aaaacagact acataatact gtaaaacaca acatatgcag tcactatgaa
      841 ccaactactt agatggtatt agtgacctgt agaattcgag ctctagagct gcagggcggc
      901 cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
      961 cccgtgtctc aaaatctctg atgttacatt gcacaagatc ggtacccggg gatcctctag
     1021 accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca
     1081 ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg
     1141 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     1201 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct
     1261 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
     1321 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg
     1381 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
     1441 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
     1501 aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
     1561 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
     1621 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
     1681 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
     1741 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1801 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1861 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1921 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1981 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     2041 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     2101 gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc
     2161 aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3001 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3061 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3121 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3181 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3241 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3301 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     3361 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcg
//
