LOCUS       sequence-263864-        4053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4053)
  AUTHORS   Superti-Furga G, Lackner D, Wiedmer T, Ingles-Prieto A, Barbosa B, 
            Girardi E, Goldmann U, Gurtl B, Klavins K, Klimek C, Lindinger S, 
            Lineiro-Retes E, Muller AC, Onstein S, Redinger G, Reil D, Sedlyarov
            V, Wolf G, Crawford M, Everley R, ... Steppan CM
  TITLE     The RESOLUTE consortium: unlocking SLC transporters for drug 
            discovery.
  JOURNAL   Nat Rev Drug Discov. 2020 Jul;19(7):429-430. doi: 
            10.1038/d41573-020-00056-6.
  PUBMED    32265506
REFERENCE   2  (bases 1 to 4053)
  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"; doi: "10"; journalName: 
            "Nat Rev Drug Discov"; date: "2020-07"; volume: "19"; issue: "7"; 
            pages: "429-430"
COMMENT     Sequence Label: pDONR221_SLC2A5
FEATURES             Location/Qualifiers
     source          1..4053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2178..2277)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2294..2313)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2295..2313)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2318..2334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2318..2334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2436..2455)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2447..3256
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2514..2533)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3403..3991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3892..3911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggagc agcaggacca gagcatgaag gagggccggc tgaccctggt
      721 gctggccctg gccacactga tcgcagcctt cggcagctca tttcaatacg ggtacaatgt
      781 cgcggccgtg aattcccctg ccctgctgat gcagcagttc tataacgaga cctactatgg
      841 ccggacaggc gagttcatgg aggattttcc actgaccctg ctgtggagcg tgacagtgtc
      901 catgttccca tttggcggct tcatcggatc cctgctggtg ggaccactgg tgaataagtt
      961 tggcagaaag ggcgccctgc tgttcaacaa tatcttttct atcgtgcctg ccatcctgat
     1021 gggctgctct agggtggcca ccagctttga gctgatcatc atcagcaggc tgctggtggg
     1081 aatctgtgca ggcgtgtcta gcaacgtggt gccaatgtat ctgggcgagc tggcaccaaa
     1141 gaatctgagg ggcgccctgg gagtggtgcc acagctgttc atcaccgtgg gcatcctggt
     1201 ggcccagatc tttggcctgc gcaacctgct ggccaatgtg gacggatggc caatcctgct
     1261 gggactgaca ggagtgcctg ccgccctgca gctgctgctg ctgccattct ttcccgagag
     1321 ccctcggtac ctgctgatcc agaagaagga cgaggcagca gccaagaagg ccctgcagac
     1381 cctgagagga tgggactccg tggataggga ggtggcagag atcagacagg aggatgaggc
     1441 agagaaggca gcaggcttca tctctgtgct gaagctgttt aggatgcgca gcctgaggtg
     1501 gcagctgctg tccatcatcg tgctgatggg aggacagcag ctgtctggag tgaacgccat
     1561 ctactattac gccgaccaga tctatctgag cgccggagtg ccagaggagc acgtgcagta
     1621 cgtgaccgca ggaacaggag cagtgaacgt ggtcatgacc ttctgcgccg tgtttgtggt
     1681 ggagctgctg ggccggagac tgctgctgct gctgggcttc tctatctgtc tgatcgcatg
     1741 ctgcgtgctg accgccgccc tggccctgca ggatacagtg agctggatgc catatatctc
     1801 catcgtgtgc gtgatctctt acgtgatcgg acacgcactg ggaccaagcc caatccctgc
     1861 cctgctgatc accgagatct tcctgcagtc ctctaggcca tccgccttta tggtgggagg
     1921 atccgtgcac tggctgtcta acttcacagt gggcctgatc ttccctttta tccaggaggg
     1981 cctgggccca tatagcttca tcgtgtttgc cgtgatctgt ctgctgacca caatctacat
     2041 ctttctgatc gtgcccgaga ccaaggccaa gacattcatc gagatcaacc agatctttac
     2101 caagatgaat aaggtgtccg aggtgtaccc tgagaaggag gagctgaagg agctgccccc
     2161 tgtgacatct gagcaggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2221 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2281 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2341 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2401 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2461 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2521 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2581 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2641 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2701 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2761 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2821 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2881 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2941 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3001 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3061 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3121 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3181 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3241 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3301 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4021 cctggccttt tgctggcctt ttgctcacat gtt
//
