LOCUS       sequence-464334-        4206 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 4206)
  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 4206)
  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-SLC22A13_STOP
FEATURES             Location/Qualifiers
     source          1..4206
                     /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(2331..2430)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2447..2466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2448..2466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2471..2487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2471..2487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2589..2608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2600..3409
                     /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(2667..2686)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3556..4144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4045..4064
                     /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 accatggccc agttcgtgca ggtgctggca gagatcggcg acttcggccg
      721 gtttcagatc cagctgctga tcctgctgtg cgtgctgaac ttcctgtctc ctttttactt
      781 ctttgcccac gtgttcatgg tgctggacga gccacaccac tgtgccgtgg cctgggtgaa
      841 gaaccacacc tttaatctgt ccgccgcaga gcagctggtg ctgtctgtgc cactggatac
      901 agcaggacac ccagagcctt gcctgatgtt caggccacct ccagcaaacg catccctgca
      961 ggacatcctg tctcaccggt tcaacgagac ccagccttgc gatatgggct gggagtaccc
     1021 agagaatagg ctgccctctc tgaagaacga gttcaacctg gtgtgcgacc gcaagcacct
     1081 gaaggatacc acacagagcg tgttcatggc cggcctgctg gtgggcaccc tgatgtttgg
     1141 ccctctgtgc gaccggatcg gcagaaaggc cacaatcctg gcccagctgc tgctgtttac
     1201 cctgatcggc ctggccacag ccttcgtgcc aagctttgag ctgtatatgg ccctgaggtt
     1261 cgcagtggca accgcagtgg caggactgag cttttccaac gtgaccctgc tgacagagtg
     1321 ggtgggccct agctggagaa cacaggccgt ggtgctggcc cagtgtaact tctccctggg
     1381 acagatggtg ctggcaggac tggcctacgg ctttaggaat tggaggctgc tgcagatcac
     1441 cggaacagca ccaggcctgc tgctgttctt ttatttctgg gcactgccag agagcgccag
     1501 gtggctgctg accaggggac gcatggatga ggccatccag ctgatccaga aggccgccag
     1561 cgtgaaccgg agaaagctgt ccccagagct gatgaatcag ctggtgcctg agaagaccgg
     1621 cccatccggc aatgccctgg acctgttcag gcacccccag ctgcgcaagg tgacactgat
     1681 catcttctgc gtgtggtttg tggattctct gggctactat ggactgagcc tgcaagtggg
     1741 cgacttcgga ctggacgtgt acctgaccca gctgatcttt ggagcagtgg aggtgccagc
     1801 aaggtgcagc tccatcttca tgatgcagcg gtttggcaga aagtggtccc agctgggcac
     1861 cctggtgctg ggaggactga tgtgcatcat catcatcttc atccctgccg atctgccagt
     1921 ggtggtgaca atgctggcag tggtgggcaa gatggcaacc gcagcagcct tcacaatctc
     1981 ttacgtgtat agcgccgagc tgtttcccac catcctgagg cagacaggaa tgggactggt
     2041 gggcatcttt tccagaatcg gcggcatcct gacccctctg gtcatcctgc tgggagagta
     2101 ccacgccgcc ctgcccatgc tgatctatgg ctctctgcct atcgtggcag gactgctgtg
     2161 caccctgctg ccagagacac acggacaggg cctgaaggac accctgcagg atctggagct
     2221 gggaccacac ccaaggtctc caaagagcgt gccttccgag aaggagaccg aggcaaaggg
     2281 acgcacatct agcccaggcg tggccttcgt gtcctctaca tatttttaag acccagcttt
     2341 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2401 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2461 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2521 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2581 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2641 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2701 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2761 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2821 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2881 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2941 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3001 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3061 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3121 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3181 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3241 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3301 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3361 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3421 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3481 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4201 catgtt
//
