LOCUS       sequence-388616-        3897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3897)
  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 3897)
  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-SLC52A1_STOP
FEATURES             Location/Qualifiers
     source          1..3897
                     /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(2022..2121)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2138..2157)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2139..2157)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2162..2178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2162..2178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2280..2299)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2291..3100
                     /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(2358..2377)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3247..3835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3736..3755
                     /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 accatggcag cacctaccct gggcaggctg gtgctgacac acctgctggt
      721 ggccctgttc ggaatgggaa gctgggcagc agtgaacgga atctgggtgg agctgccagt
      781 ggtggtgaag gacctgcctg agggctggtc cctgccatct tacctgagcg tggtggtggc
      841 cctgggaaat ctgggactgc tggtggtgac cctgtggagg cagctggcac caggcaaggg
      901 agagcaggtg cctatccagg tggtgcaggt gctgtccgtg gtgggcaccg ccctgctggc
      961 cccactgtgg caccacgtgg caccagtggc aggacagctg cactctgtgg ccttcctgac
     1021 actggccctg gtgctggcca tggcctgctg tacctctaac gtgacattcc tgccatttct
     1081 gagccacctg ccccctccat ttctgcggtc tttctttctg ggacagggac tgagcgccct
     1141 gctgccatgc gtgctggccc tggtgcaggg agtgggccgg ctggagtgtc cccctgcccc
     1201 caccaatggc acatctggcc cacccctgga cttcccagag agatttcccg ccagcacctt
     1261 cttttgggcc ctgaccgccc tgctggtgac atccgccgca gccttccggg gactgctgct
     1321 gctgctgcct tccctgccat ctgtgaccac aggaggatcc ggacctgagc tgcagctggg
     1381 atctccagga gcagaggagg aggagaagga ggaggaggag gccctgcctc tgcaggagcc
     1441 tccatctcag gcagcaggaa caatccctgg accagatcca gaggcacacc agctgttcag
     1501 cgcccacgga gcctttctgc tgggactgat ggccttcacc tccgccgtga caaacggcgt
     1561 gctgccctct gtgcagtctt ttagctgcct gccttacggc aggctggcct atcacctggc
     1621 agtggtgctg ggaagcgccg caaatccact ggcatgcttc ctggcaatgg gcgtgctgtg
     1681 ccgcagcctg gcaggactgg tgggcctgtc cctgctgggc atgctgtttg gcgcctacct
     1741 gatggcactg gcaatcctgt ccccttgtcc acctctggtg ggaaccacag caggagtggt
     1801 gctggtggtg ctgagctggg tgctgtgcct gtgcgtgttc agctatgtga aggtggccgc
     1861 cagctccctg ctgcacggag gaggccggcc cgccctgctg gcagccggcg tggccatcca
     1921 agtgggctcc ctgctgggcg ccggcgccat gttcccaccc accagcatct atcacgtgtt
     1981 tcagtccaga aaggactgcg tggatccctg tggcccttaa gacccagctt tcttgtacaa
     2041 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2101 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2161 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2221 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2281 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2341 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2401 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2461 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2521 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2581 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2641 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2701 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2761 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2821 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2881 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2941 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3001 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3061 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3121 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3181 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3241 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3301 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3361 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3421 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3481 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3541 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3601 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3661 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3721 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3781 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3841 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
