LOCUS       sequence-259497-        4428 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 4428)
  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 4428)
  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_SLC13A4
FEATURES             Location/Qualifiers
     source          1..4428
                     /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(2553..2652)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2669..2688)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2670..2688)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2693..2709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2693..2709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2811..2830)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2822..3631
                     /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(2889..2908)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3778..4366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4267..4286
                     /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 accatgggac tgctgcaggg actgctgcgg gtgagaaagc tgctgctggt
      721 ggtgtgcgtg ccactgctgc tgctgcctct gccagtgctg caccctagct ccgaggcctc
      781 ttgtgcctac gtgctgatcg tgaccgccgt gtattgggtg agcgaggcag tgcctctggg
      841 agcagccgcc ctggtgcccg cctttctgta ccctttcttt ggcgtgctga ggagcaatga
      901 ggtggccgcc gagtatttca agaacaccac actgctgctg gtgggcgtga tctgcgtggc
      961 agcagccgtg gagaagtgga atctgcacaa gcggatcgcc ctgagaatgg tgctgatggc
     1021 aggagcaaag ccaggaatgc tgctgctgtg ctttatgtgc tgtaccacac tgctgtctat
     1081 gtggctgagc aacacctcca ccacagccat ggtcatgcct atcgtggagg ccgtgctgca
     1141 ggagctggtg tccgccgagg acgagcagct ggtggcaggc aactctaata cagaggaggc
     1201 cgagcctatc agcctggatg tgaagaattc tcagccaagc ctggagctga tcttcgtgaa
     1261 tgaggagagc aacgccgacc tgaccacact gatgcacaac gagaatctga acggcgtgcc
     1321 atccatcacc aatcccatca agacagccaa ccagcaccag ggcaagaagc agcaccctag
     1381 ccaggagaag ccacaggtgc tgaccccctc ccctcgcaag cagaagctga acaggaagta
     1441 ccgcagccac cacgatcaga tgatctgcaa gtgtctgagc ctgtccatct cttatagcgc
     1501 cacaatcggc ggcctgacca caatcatcgg cacctccaca tctctgatct ttctggagca
     1561 cttcaacaat cagtaccctg ccgccgaggt ggtgaatttt ggcacctggt ttctgttctc
     1621 ctttccaatc tctctgatca tgctggtggt gtcctggttc tggatgcact ggctgtttct
     1681 gggctgcaac ttcaaggaga cctgtagcct gagcaagaag aagaagacca agagggagca
     1741 gctgtctgag aagcgcatcc aggaggagta cgagaagctg ggcgacatca gctatcccga
     1801 gatggtgacc ggcttctttt tcatcctgat gaccgtgctg tggtttacaa gggagccagg
     1861 attcgtgcca ggatgggact cctttttcga gaagaagggc tacagaaccg atgccaccgt
     1921 gagcgtgttc ctgggctttc tgctgttcct gatccctgcc aagaagccat gctttggcaa
     1981 gaagaatgac ggcgagaacc aggagcacag cctgggcacc gagccaatca tcacatggaa
     2041 ggatttccag aagaccatgc cctgggagat cgtgatcctg gtgggaggag gatatgcact
     2101 ggcatctggc agcaagtcta gcggcctgtc cacatggatc ggcaatcaga tgctgtctct
     2161 gtcctctctg ccaccttggg cagtgaccct gctggcatgc atcctggtga gcatcgtgac
     2221 agagttcgtg agcaatccag ccaccatcac aatcttcctg cccatcctgt gctccctgtc
     2281 tgagaccctg cacatcaacc ccctgtacac cctgatccct gtgacaatgt gcatctcctt
     2341 tgccgtgatg ctgccagtgg gcaatccacc caacgccatc gtgttctctt atggccactg
     2401 tcagatcaag gatatggtga aggccggcct gggcgtgaat gtgatcggcc tggtcatcgt
     2461 gatggtggcc atcaacacct ggggcgtgag cctgttccac ctggacacat atccagcatg
     2521 ggcacgggtg tccaacatca ccgatcaggc cgacccagct ttcttgtaca aagttggcat
     2581 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2641 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2701 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2761 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2821 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2881 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2941 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3001 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3061 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3121 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3181 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3241 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3301 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3361 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3421 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3481 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3541 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3601 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3661 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3721 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3781 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3841 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3901 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3961 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4021 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4081 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4141 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4201 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4261 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4321 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4381 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
