LOCUS       sequence-259443-        4326 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 4326)
  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 4326)
  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_SLC13A2
FEATURES             Location/Qualifiers
     source          1..4326
                     /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(2451..2550)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2567..2586)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2568..2586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2591..2607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2591..2607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2709..2728)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2720..3529
                     /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(2787..2806)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3676..4264
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4165..4184
                     /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 accatggcaa cctgctggca ggccctgtgg gcataccggt cttatctgat
      721 cgtgttcttt gtgcctatcc tgctgctgcc actgcccatc ctggtgccaa gcaaggaggc
      781 ctactgcgcc tatgccatca tcctgatggc cctgttctgg tgtacagagg ccctgccact
      841 ggcagtgacc gccctgtttc ctctgatcct gttcccaatg atgggaatcg tggacgcatc
      901 cgaggtggca gtggagtacc tgaaggattc taacctgctg ttctttggag gactgctggt
      961 ggcaatcgca gtggagcact ggaatctgca caagcggatc gccctgagag tgctgctgat
     1021 cgtgggcgtg cggcctgcac cactgatcct gggctttatg ctggtgaccg ccttcctgag
     1081 catgtggatc tccaacaccg ccacatccgc catgatggtg ccaattgctc acgctgttct
     1141 ggatcagctg cacagctccc aggcctctag caatgtggag gagggctcta acaatcccac
     1201 cttcgagctg caggagccca gccctcagaa ggaggtgaca aagctggata acggacaggc
     1261 cctgcctgtg accagcgcct cctctgaggg aagagcacac ctgtcccaga agcacctgca
     1321 cctgacacag tgcatgtccc tgtgcgtgtg ctacagcgcc tccatcggag gaatcgccac
     1381 cctgacagga accgcaccaa acctggtgct gcagggacag atcaattctc tgttccccca
     1441 gaacggcaat gtggtgaact ttgcctcttg gttcagcttt gccttcccta ccatggtcat
     1501 cctgctgctg ctggcctggc tgtggctgca gatcctgttc ctgggcttta acttcaggaa
     1561 gaattttggc atcggcgaga agatgcagga gcagcagcag gccgcctatt gcgtgatcca
     1621 gacagagcac cgcctgctgg gcccaatgac cttcgccgag aaggccatct ccatcctgtt
     1681 cgtgatcctg gtgctgctgt ggtttacaag ggagcccggc ttctttctgg gctggggcaa
     1741 cctggccttt cctaatgcca agggcgagtc catggtgtct gacggcacag tggccatctt
     1801 tatcggcatc atcatgttca tcatcccaag caagtttccc ggcctgaccc aggaccccga
     1861 gaatcctggc aagctgaagg cccctctggg cctgctggat tggaagacag tgaaccagaa
     1921 gatgccatgg aatatcgtgc tgctgctggg aggaggatac gcactggcca agggaagcga
     1981 gcggagcgga ctgtctgagt ggctgggcaa taagctgacc ccactgcagt ccgtgccagc
     2041 accagcaatc gcaatcatcc tgtctctgct ggtggccaca ttcaccgagt gcacaagcaa
     2101 cgtggccacc acaaccatct ttctgccaat cctggccagc atggcacagg caatctgcct
     2161 gcacccactg tatgtgatgc tgccttgtac actggccacc tccctggcct tcatgctgcc
     2221 agtggcaacc ccacctaacg ccatcgtgtt ttctttcggc gacctgaagg tgctggatat
     2281 ggcccgggcc ggctttctgc tgaacatcat cggcgtgctg atcatcgccc tggccatcaa
     2341 tagctggggc atcccactgt ttagcctgca ctccttcccc tcttgggccc agtccaacac
     2401 aaccgcccag tgtctgccct ctctggccaa tacaaccaca cctagcccag acccagcttt
     2461 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2521 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2581 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2641 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2701 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2761 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2821 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2881 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2941 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3001 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3061 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3121 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3181 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3241 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3301 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3361 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3421 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3481 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3541 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3601 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3661 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3721 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3781 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3841 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3901 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3961 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4021 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4081 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4141 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4201 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4261 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4321 catgtt
//
