LOCUS       sequence-312132-        4431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4431)
  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 4431)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4431
                     /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(2556..2655)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2672..2691)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2673..2691)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2696..2712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2696..2712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2814..2833)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2825..3634
                     /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(2892..2911)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3781..4369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4270..4289
                     /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 ctaagaccca gctttcttgt acaaagttgg
     2581 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2641 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2701 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2761 taaaataata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2821 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2881 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2941 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     3001 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     3061 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     3121 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     3181 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     3241 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     3301 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3361 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3421 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3481 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3541 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3601 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3661 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3721 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3781 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3841 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3901 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3961 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4021 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4081 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4141 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4201 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4261 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4321 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4381 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
