LOCUS       sequence-261014-        4506 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 4506)
  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 4506)
  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_SLC20A2
FEATURES             Location/Qualifiers
     source          1..4506
                     /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(2631..2730)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2747..2766)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2748..2766)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2771..2787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2771..2787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2889..2908)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2900..3709
                     /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(2967..2986)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3856..4444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4345..4364
                     /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 tggacgagta tctgtggatg gtcatcctgg gcttcatcat
      721 cgcctttatc ctggccttca gcgtgggcgc caacgatgtg gccaattcct ttggcaccgc
      781 agtgggatct ggagtggtga cactgaggca ggcctgcatc ctggccagca tcttcgagac
      841 cacaggatcc gtgctgctgg gagcaaaagt gggagagacc atccgcaagg gcatcatcga
      901 cgtgaacctg tacaatgaga ccgtggagac actgatggca ggagaggtga gcgccatggt
      961 gggatccgcc gtgtggcagc tgatcgcatc ttttctgcgg ctgcccatca gcggcaccca
     1021 ctgtatcgtg ggctctacaa tcggcttcag cctggtggcc atcggcacaa agggcgtgca
     1081 gtggatggag ctggtgaaga tcgtggcctc ttggttcatc agccctctgc tgtccggctt
     1141 catgtctggc ctgctgtttg tgctgatccg gatcttcatc ctgaagaagg aggacccagt
     1201 gcctaacgga ctgagagccc tgccagtgtt ttatgccgcc accatcgcca tcaacgtgtt
     1261 cagcatcatg tacacaggag cacctgtgct gggactggtg ctgccaatgt gggccatcgc
     1321 cctgatctct tttggcgtgg ccctgctgtt cgccttcttc gtgtggctgt tcgtgtgccc
     1381 ctggatgcgg agaaagatca ccggcaagct gcagaaggag ggcgccctgt ccagggtgtc
     1441 tgatgagagc ctgtccaagg tgcaggaggc cgagtctcca gtgttcaagg agctgccagg
     1501 agcaaaggca aacgacgata gcaccatccc tctgacagga gcagcaggag agaccctggg
     1561 cacatccgag ggcaccagcg ccggatccca ccctagggca gcatacggaa gagccctgag
     1621 catgacacac ggctctgtga agagcccaat ctccaatggc accttcggct ttgacggcca
     1681 cacacgcagc gatggccacg tgtatcacac cgtgcacaag gactccggcc tgtacaagga
     1741 tctgctgcac aagatccaca tcgacagggg accagaggag aagcctgcac aggagtccaa
     1801 ctatcgcctg ctgaggcgca acaattctta cacctgctat acagcagcaa tctgtggact
     1861 gccagtgcac gccaccttcc gggcagcaga cagctccgcc ccagaggatt ctgagaagct
     1921 ggtgggcgac acagtgtctt acagcaagaa gaggctgcgc tacgatagct attctagcta
     1981 ctgcaatgca gtggcagagg cagagatcga ggcagaggag ggcggcgtgg agatgaagct
     2041 ggcatccgag ctggcagacc cagatcagcc aagagaggac ccagcagagg aggagaagga
     2101 ggagaaggat gcccctgagg tgcacctgct gttccacttt ctgcaggtgc tgaccgcatg
     2161 cttcggaagc tttgcccacg gaggcaacga cgtgagcaat gcaatcggac cactggtggc
     2221 cctgtggctg atctataagc agggaggagt cacccaggag gcagcaacac ccgtgtggct
     2281 gctgttttac ggaggagtgg gaatctgtac cggactgtgg gtgtggggcc ggagagtgat
     2341 ccagacaatg ggcaaggacc tgacccctat cacaccatcc tctggcttta ccatcgagct
     2401 ggccagcgcc ttcacagtgg tcatcgcctc caacatcggc ctgcccgtgt ctaccacaca
     2461 ctgtaaagtg ggatctgtgg tggcagtggg atggatccgg agcagaaagg ccgtggattg
     2521 gaggctgttt cgcaatatct tcgtggcatg gtttgtgacc gtgccagtgg caggactgtt
     2581 ctccgccgcc gtgatggccc tgctgatgta tggcatcctg ccttacgtgg acccagcttt
     2641 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2701 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2761 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2821 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2881 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2941 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3001 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3061 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3121 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3181 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3241 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3301 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3361 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3421 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3481 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3541 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3601 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3661 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3721 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3781 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3841 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3901 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3961 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4021 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4081 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4141 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4201 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4261 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4321 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4381 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4441 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4501 catgtt
//
