LOCUS       sequence-162640-        3738 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway Donor vector containing AKR1B1, used as an over expression 
            control for gene expression analysis. Part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3738)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 3738)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_AKR1B1_WT_V5
FEATURES             Location/Qualifiers
     source          1..3738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1138..1237)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1254..1273)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1255..1273)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1278..1294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1278..1294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1725..2516
                     /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      2609..3197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3098..3117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3351..3368
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3527..3554)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3646..3732)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcat ggcaagccgt ctcctgctca acaacggcgc caagatgccc atcctggggt
      241 tgggtacctg gaagtcccct ccagggcagg tgactgaggc cgtgaaggtg gccattgacg
      301 tcgggtaccg ccacatcgac tgtgcccatg tgtaccagaa tgagaatgag gtgggggtgg
      361 ccattcagga gaagctcagg gagcaggtgg tgaagcgtga ggagctcttc atcgtcagca
      421 agctgtggtg cacgtaccat gagaagggcc tggtgaaagg agcctgccag aagacactca
      481 gcgacctgaa gctggactac ctggacctct accttattca ctggccgact ggctttaagc
      541 ctgggaagga atttttccca ttggatgagt cgggcaatgt ggttcccagt gacaccaaca
      601 ttctggacac gtgggcggcc atggaagagc tggtggatga agggctggtg aaagctattg
      661 gcatctccaa cttcaaccat ctccaggtgg agatgatctt aaacaaacct ggcttgaagt
      721 ataagcctgc agttaaccag attgagtgcc acccatatct cactcaggag aagttaatcc
      781 agtactgcca gtccaaaggc atcgtggtga ccgcctacag ccccctcggc tctcctgaca
      841 ggccctgggc caagcccgag gacccttctc tcctggagga tcccaggatc aaggcgatcg
      901 cagccaagca caataaaact acagcccagg tcctgatccg gttccccatg cagaggaact
      961 tggtggtgat ccccaagtct gtgacaccag aacgcattgc tgagaacttt aaggtctttg
     1021 actttgaact gagcagccag gatatgacca ccttactcag ctacaacagg aactggaggg
     1081 tctgtgcctt gttgagctgt acctcccaca aggattaccc cttccatgaa gagttttgcc
     1141 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     1201 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1261 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1321 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     1381 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     1441 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     1501 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     1561 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     1621 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     1681 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     1741 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     1801 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     1861 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     1921 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     1981 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2041 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2101 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     2161 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     2221 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     2281 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     2341 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     2401 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     2461 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     2521 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     2581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     2761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     2821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3241 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3301 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3361 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3421 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     3481 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     3541 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     3601 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     3661 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     3721 tcgttttatt tgatgcct
//
