LOCUS       sequence-260960-        4587 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 4587)
  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 4587)
  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_SLC20A1
FEATURES             Location/Qualifiers
     source          1..4587
                     /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(2712..2811)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2828..2847)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2829..2847)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2852..2868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2852..2868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2970..2989)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2981..3790
                     /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(3048..3067)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3937..4525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4426..4445
                     /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 ccctgatcac atctaccaca gcagcaaccg cagcaagcgg
      721 accactggtg gactacctgt ggatgctgat cctgggcttc atcatcgcct ttgtgctggc
      781 cttcagcgtg ggcgccaacg atgtggccaa tagcttcggc accgcagtgg gatccggagt
      841 ggtgacactg aagcaggcct gcatcctggc ctctatcttt gagaccgtgg gaagcgtgct
      901 gctgggagca aaggtgtccg agacaatccg gaagggcctg atcgacgtgg agatgtataa
      961 ctctacacag ggactgctga tggcaggaag cgtgtccgcc atgttcggat ccgccgtgtg
     1021 gcagctggtg gcatcctttc tgaagctgcc tatctctggc acccactgta tcgtgggcgc
     1081 cacaatcggc ttctctctgg tggccaaggg ccaggagggc gtgaagtgga gcgagctgat
     1141 caagatcgtg atgtcttggt tcgtgagccc tctgctgtcc ggcatcatgt ctggcatcct
     1201 gttctttctg gtgagggcct ttatcctgca caaggcagac ccagtgccta acggactgcg
     1261 cgccctgcca gtgttctacg cctgcaccgt gggcatcaat ctgtttagca tcatgtatac
     1321 aggcgcccct ctgctgggct tcgataagct gccactgtgg ggcaccatcc tgatctccgt
     1381 gggctgcgcc gtgttttgtg ccctgatcgt gtggttcttc gtgtgcccac ggatgaagag
     1441 aaagatcgag cgggagatca agtgttctcc tagcgagtcc ccactgatgg agaagaagaa
     1501 cagcctgaag gaggaccacg aggagacaaa gctgtccgtg ggcgatatcg agaataagca
     1561 ccccgtgtct gaagtgggac cagcaaccgt gcctctgcag gccgtggtgg aggagagaac
     1621 agtgagcttc aagctgggcg acctggagga ggcaccagag agggagagac tgccatccgt
     1681 ggacctgaag gaggagacct ccatcgattc tacagtgaac ggagcagtgc agctgccaaa
     1741 cggaaatctg gtgcagttta gccaggccgt gtccaaccag atcaatagct ccggccacta
     1801 ccagtatcac accgtgcaca aggactctgg cctgtacaag gagctgctgc acaagctgca
     1861 cctggccaaa gtgggcgatt gcatgggcga ctccggcgat aagccactga ggagaaacaa
     1921 tagctacacc tcctatacaa tggccatctg tggcatgcct ctggatagct tcagggcaaa
     1981 ggagggagag cagaagggag aggagatgga gaagctgacc tggcctaacg ccgactccaa
     2041 gaagaggatc cgcatggatt cttacacaag ctattgcaat gccgtgagcg acctgcactc
     2101 tgccagcgag atcgatatgt ccgtgaaggc agagatggga ctgggcgaca ggaagggatc
     2161 caatggctct ctggaggagt ggtacgacca ggataagcca gaggtgagcc tgctgttcca
     2221 gtttctgcag atcctgaccg catgcttcgg atcctttgcc cacggaggca acgacgtgag
     2281 caatgccatc ggcccactgg tggccctgta cctggtgtat gacaccggcg acgtgagcag
     2341 caaggtggcc acacccatct ggctgctgct gtatggagga gtgggaatct gcgtgggact
     2401 gtgggtgtgg ggcaggcgcg tgatccagac catgggcaag gacctgaccc caatcacacc
     2461 ctcctctggc ttctccatcg agctggcctc tgccctgaca gtggtcatcg ccagcaacat
     2521 cggcctgccc atctccacca cacactgtaa agtgggaagc gtggtgtccg tgggatggct
     2581 gcggagcaag aaggcagtgg attggaggct gtttcgcaat atcttcatgg cctggtttgt
     2641 gaccgtgcct atctctggcg tgatcagcgc cgccatcatg gccatctttc ggtacgtgat
     2701 cctgagaatg gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2761 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2821 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2881 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2941 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3001 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3061 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3121 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3181 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3241 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3301 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3361 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3421 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3481 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3541 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3601 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3661 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3721 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3781 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3841 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3901 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4081 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4501 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4561 cttttgctgg ccttttgctc acatgtt
//
