LOCUS       sequence-259541-        4035 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 4035)
  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 4035)
  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_SLC17A5
FEATURES             Location/Qualifiers
     source          1..4035
                     /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(2160..2259)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2276..2295)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2277..2295)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2300..2316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2300..2316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2418..2437)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2429..3238
                     /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(2496..2515)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3385..3973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3874..3893
                     /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 accatgagga gccccgtgcg cgacctggca aggaacgatg gcgaggagtc
      721 caccgacaga acaccactgc tgccaggagc acctagggca gaggcagcac ccgtgtgctg
      781 tagcgccaga tacaacctgg ccatcctggc cttctttggc ttctttatcg tgtatgccct
      841 gagggtgaat ctgagcgtgg ccctggtgga catggtggat tccaacacca cactggagga
      901 caatcgcacc tctaaggcct gccctgagca cagcgcccca atcaaggtgc accacaatca
      961 gacaggcaag aagtaccagt gggatgccga gacccagggc tggatcctgg gctctttctt
     1021 ttacggctat atcatcacac agatcccagg cggctatgtg gcctccaaga tcggcggcaa
     1081 gatgctgctg ggattcggaa tcctgggaac cgccgtgctg accctgttta caccaatcgc
     1141 cgccgacctg ggcgtgggcc ccctgatcgt gctgcgcgcc ctggagggac tgggagaggg
     1201 agtgacattc cctgccatgc acgcaatgtg gagctcctgg gcaccacctc tggagcggag
     1261 caagctgctg tccatctctt acgcaggagc acagctggga accgtgatca gcctgccact
     1321 gtccggcatc atctgttact atatgaactg gacatacgtg ttttatttct ttggcaccat
     1381 cggcatcttc tggtttctgc tgtggatctg gctggtgtcc gatacccccc agaagcacaa
     1441 gcgcatctct cactacgaga aggagtatat cctgtctagc ctgcggaacc agctgtcctc
     1501 tcagaagtct gtgccatggg tgcccatcct gaagagcctg cctctgtggg caatcgtggt
     1561 ggcacacttc tcctacaatt ggacctttta tacactgctg accctgctgc ctacctacat
     1621 gaaggagatc ctgagattca acgtgcagga gaatggcttt ctgagctccc tgccatatct
     1681 gggctcctgg ctgtgcatga tcctgtctgg ccaggccgcc gataacctga gggccaagtg
     1741 gaacttcagc accctgtgcg tgcggagaat cttttctctg atcggcatga tcggaccagc
     1801 cgtgttcctg gtggcagcag gctttatcgg ctgcgactac agcctggccg tggccttcct
     1861 gaccatctcc accacactgg gcggcttctg ttctagcggc ttttctatca accacctgga
     1921 tatcgcccca agctatgccg gcatcctgct gggcatcacc aatacattcg ccacaatccc
     1981 cggcatggtg ggccctgtga tcgccaagag cctgacaccc gacaacaccg tgggcgagtg
     2041 gcagacagtg ttttacatcg ccgccgccat caacgtgttc ggcgccatct tctttaccct
     2101 gtttgccaag ggcgaggtgc agaactgggc cctgaatgat caccacggcc accggcacga
     2161 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2221 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2281 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2341 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2401 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2461 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2521 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2581 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2641 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2701 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2761 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2821 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2881 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2941 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3001 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3061 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3121 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3181 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3241 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3301 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4021 ttttgctcac atgtt
//
