LOCUS       sequence-262916-        4356 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 4356)
  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 4356)
  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_SLC13A3
FEATURES             Location/Qualifiers
     source          1..4356
                     /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(2481..2580)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2597..2616)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2598..2616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2621..2637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2621..2637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2739..2758)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2750..3559
                     /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(2817..2836)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3706..4294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4195..4214
                     /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 accatggccg ccctggcagc agcagccaag aaggtctggt ccgcccggag
      721 actgctggtg ctgctgttta ccccactggc cctgctgcca gtggtgttcg cactgccacc
      781 taaggagggc aggtgcctgt ttgtgatcct gctgatggcc gtgtactggt gtaccgaggc
      841 cctgccactg tccgtgacag ccctgctgcc tatcgtgctg tttccattca tgggcatcct
      901 gccctctaac aaggtgtgcc ctcagtattt cctggacacc aattttctgt tcctgagcgg
      961 cctgatcatg gcctccgcca tcgaggagtg gaacctgcac aggcgcatcg ccctgaagat
     1021 cctgatgctg gtgggagtgc agcctgcaag gctgatcctg ggcatgatgg tgaccacatc
     1081 ttttctgagc atgtggctgt ctaacaccgc cagcacagcc atgatgctgc ctatcgccaa
     1141 tgccatcctg aagtccctgt tcggccagaa ggaggtgcgc aaggatccat cccaggagtc
     1201 tgaggagaac accgccgccg tgcggagaaa tggcctgcac accgtgccaa cagagatgca
     1261 gtttctggca agcaccgagg caaaggacca cccaggagag acagaggtgc cactggacct
     1321 gcccgccgat tctcggaagg aggatgagta caggcgcaat atctggaagg gcttcctgat
     1381 cagcatccca tatagcgcct ccatcggagg aaccgccaca ctgaccggaa cagcaccaaa
     1441 cctgatcctg ctgggccagc tgaagagctt ctttcctcag tgcgacgtgg tgaattttgg
     1501 ctcctggttt atcttcgcct ttcctctgat gctgctgttc ctgctggccg gctggctgtg
     1561 gatctctttt ctgtacggag gactgagctt ccggggatgg agaaagaaca agagcgagat
     1621 caggaccaat gcagaggata gggcaagagc cgtgatcagg gaggagtatc agaacctggg
     1681 ccctatcaag ttcgccgagc aggccgtgtt catcctgttt tgtatgttcg ccatcctgct
     1741 gtttacccgg gaccctaagt tcatcccagg ctgggcctcc ctgtttaatc caggcttcct
     1801 gtctgatgcc gtgaccggcg tggccatcgt gacaatcctg ttctttttcc cctcccagag
     1861 accttctctg aagtggtggt tcgactttaa ggccccaaac accgagacag agcccctgct
     1921 gacctggaag aaggcccagg agacagtgcc atggaatatc atcctgctgc tgggaggagg
     1981 attcgcaatg gcaaagggat gcgaggagag cggactgtcc gtgtggatcg gaggacagct
     2041 gcacccactg gagaacgtgc cacccgccct ggccgtgctg ctgatcaccg tggtcatcgc
     2101 ctttttcaca gagtttgcct ctaatactgc cacaatcatc atcttcctgc ccgtgctggc
     2161 agagctggca atcaggctgc gcgtgcaccc tctgtacctg atgatcccag gcaccgtggg
     2221 ctgtagcttc gcctttatgc tgcctgtgag cacacctcca aactccatcg cctttgcctc
     2281 tggccacctg ctggtgaagg acatggtgcg gaccggcctg ctgatgaacc tgatgggcgt
     2341 gctgctgctg agcctggcca tgaatacctg ggcccagaca atctttcagc tgggcacatt
     2401 ccccgactgg gccgatatgt attccgtgaa cgtgaccgcc ctgcccccta cactggccaa
     2461 tgataccttc agaacactgg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2521 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2581 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2641 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     2701 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2761 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2821 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2881 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2941 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3001 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3061 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3121 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3181 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3241 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3301 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3361 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3421 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3481 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3541 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3601 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3661 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3721 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3781 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3841 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3901 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3961 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4021 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4081 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4141 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4201 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4261 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4321 gttcctggcc ttttgctggc cttttgctca catgtt
//
