LOCUS       sequence-265763-        3966 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 3966)
  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 3966)
  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_SLC38A5
FEATURES             Location/Qualifiers
     source          1..3966
                     /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(2091..2190)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2207..2226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2208..2226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2231..2247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2231..2247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2349..2368)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2360..3169
                     /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(2427..2446)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3316..3904
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3805..3824
                     /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 tgcaggaccc caagatgaac ggcgccctgc ctagcgatgc
      721 agtgggatac aggcaggaga gagagggctt cctgccatct aggggaccag cacctggcag
      781 caagcctgtg cagttcatgg actttgaggg caagacctcc ttcggcatga gcgtgttcaa
      841 cctgagcaat gccatcatgg gatccggaat cctgggactg gcatatgcaa tggcacacac
      901 cggcgtgatc ttctttctgg ccctgctgct gtgcatcgcc ctgctgagct cctactccat
      961 ccacctgctg ctgacatgtg ccggcatcgc cggcatcagg gcatatgagc agctgggaca
     1021 gagggccttc ggccctgcag gcaaggtggt ggtggccacc gtgatctgcc tgcacaacgt
     1081 gggcgccatg tctagctacc tgtttatcat caagagcgag ctgccactgg tcatcggcac
     1141 attcctgtat atggaccccg agggcgattg gtttctgaag ggcaatctgc tgatcatcat
     1201 cgtgtccgtg ctgatcatcc tgcccctggc cctgatgaag cacctgggct acctgggcta
     1261 tacctccggc ctgtctctga catgcatgct gttctttctg gtgagcgtga tctacaagaa
     1321 gttccagctg ggctgtgcca tcggccacaa cgagacagcc atggagtctg aggccctggt
     1381 gggactgcct agccagggcc tgaattcctc ttgtgaggcc cagatgttta ccgtggatag
     1441 ccagatgtcc tacacagtgc ctatcatggc cttcgccttc gtgtgccacc cagaggtgct
     1501 gcccatctat accgagctgt gccggccatc caagcggaga atgcaggcag tggcaaacgt
     1561 gagcatcgga gccatgttct gcatgtacgg cctgaccgcc acattcggct acctgacctt
     1621 ctacagctcc gtgaaggccg agatgctgca catgtattct cagaaggacc cactgatcct
     1681 gtgcgtgcgg ctggccgtgc tgctggccgt gaccctgaca gtgcctgtgg tgctgtttcc
     1741 aatcaggcgc gccctgcagc agctgctgtt cccaggcaag gccttttcct ggccaaggca
     1801 cgtggcaatc gccctgatcc tgctggtgct ggtgaacgtg ctggtcatct gcgtgcccac
     1861 catcagagat atcttcggcg tgatcggctc tacaagcgcc ccttccctga tcttcatcct
     1921 gccatctatc ttttacctga ggatcgtgcc cagcgaggtg gagccttttc tgtcctggcc
     1981 aaagatccag gccctgtgct tcggcgtgct gggcgtgctg tttatggccg tgagcctggg
     2041 cttcatgttt gccaattggg ccaccggcca gtctcgcatg agcggccacg acccagcttt
     2101 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2161 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2221 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2281 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2341 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2401 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2461 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2521 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2581 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2641 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2701 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2761 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2821 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2881 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2941 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3001 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3061 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3121 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3181 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3241 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3301 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3361 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3421 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3481 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3541 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3601 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3661 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3721 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3781 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3841 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3901 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3961 catgtt
//
