LOCUS       sequence-259054-        4170 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 4170)
  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 4170)
  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_SLC2A9
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(2295..2394)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2411..2430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2412..2430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2553..2572)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2564..3373
                     /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(2631..2650)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3520..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4009..4028
                     /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 accatggcca ggaagcagaa ccgcaatagc aaggagctgg gactggtgcc
      721 actgaccgac gatacatccc acgcaggacc acctggacct ggaagggccc tgctggagtg
      781 cgaccacctg cgcagcggag tgcctggagg ccggagaagg aaggattggt cttgtagcct
      841 gctggtggca tccctggcag gagccttcgg cagctccttt ctctatggat acaacctgag
      901 tgtggtcaat gcccccaccc cttacatcaa ggccttctat aacgagagct gggagcgccg
      961 gcacggcagg ccaatcgacc ccgataccct gacactgctg tggtctgtga ccgtgagcat
     1021 ctttgccatc ggcggcctgg tgggcacact gatcgtgaag atgatcggca aggtgctggg
     1081 aaggaagcac accctgctgg ccaacaatgg cttcgccatc tctgccgccc tgctgatggc
     1141 atgcagcctg caggcaggag ccttcgagat gctgatcgtg ggccggttta tcatgggcat
     1201 cgacggagga gtggccctgt ccgtgctgcc aatgtacctg tccgagatct ctcccaagga
     1261 gatcagaggc tctctgggcc aggtgaccgc catcttcatc tgtatcggcg tgtttacagg
     1321 acagctgctg ggactgccag agctgctggg caaggagtct acctggcctt acctgttcgg
     1381 cgtgatcgtg gtgccagcag tggtgcagct gctgagcctg ccttttctgc cagactcccc
     1441 ccggtatctg ctgctggaga agcacaatga ggccagagcc gtgaaggcct tccagacatt
     1501 tctgggcaag gccgacgtga gccaggaggt ggaggaggtg ctggcagaga gcagggtgca
     1561 gcggtccatc agactggtgt ctgtgctgga gctgctgagg gcaccatacg tgagatggca
     1621 ggtggtgacc gtgatcgtga caatggcctg ctaccagctg tgcggcctga acgccatctg
     1681 gttctatacc aatagcatct ttggcaaggc cggcatccca cccgccaaga tcccttatgt
     1741 gaccctgtcc acaggaggaa tcgagacact ggcagccgtg ttctctggcc tggtcatcga
     1801 gcacctgggc agaaggccac tgctgatcgg cggctttggc ctgatgggcc tgttctttgg
     1861 caccctgaca atcaccctga cactgcagga ccacgcccct tgggtgccat acctgagcat
     1921 cgtgggcatc ctggccatca tcgcaagctt ctgctccgga ccaggaggca tcccttttat
     1981 cctgaccggc gagttctttc agcagtccca gaggccagca gccttcatca tcgcaggaac
     2041 agtgaactgg ctgtctaatt ttgccgtggg cctgctgttc ccctttatcc agaagagcct
     2101 ggatacctat tgcttcctgg tgtttgccac catctgtatc acaggcgcca tctacctgta
     2161 tttcgtgctg cccgagacca agaacaggac atacgccgag atctcccagg ccttttctaa
     2221 gcgcaataag gcctatcctc cagaggagaa gatcgactcc gccgtgacag atggcaagat
     2281 caacggcaga cctgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2341 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2401 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2461 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2521 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2581 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2641 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2701 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2761 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2821 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2881 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2941 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3001 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3061 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3121 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3181 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3241 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3301 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3361 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3421 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt
//
