LOCUS       sequence-312204-        3498 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 3498)
  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 3498)
  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-SLC25A31_STOP
FEATURES             Location/Qualifiers
     source          1..3498
                     /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(1623..1722)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1739..1758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1740..1758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1763..1779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1763..1779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1881..1900)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1892..2701
                     /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(1959..1978)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2848..3436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3337..3356
                     /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 accatgcacc gcgagccagc caagaagaag gccgagaagc ggctgttcga
      721 cgccagctcc tttggcaagg atctgctggc aggaggagtg gcagcagccg tgagcaagac
      781 cgccgtggcc cccatcgaga gagtgaagct gctgctgcag gtgcaggcct ctagcaagca
      841 gatcagccca gaggccaggt ataagggcat ggtggactgc ctggtgagaa tccccaggga
      901 gcagggcttc ttttccttct ggcgcggcaa cctggccaat gtgatccggt attttcctac
      961 acaggccctg aacttcgcct ttaaggataa gtacaagcag ctgttcatgt ccggcgtgaa
     1021 caaggagaag cagttctgga gatggtttct ggccaatctg gcatctggag gagcagcagg
     1081 agccaccagc ctgtgcgtgg tgtatcctct ggacttcgca aggacaaggc tgggagtgga
     1141 tatcggcaag ggcccagagg agcgccagtt taagggcctg ggcgactgta tcatgaagat
     1201 cgccaagtct gatggaatcg caggactgta ccagggattc ggcgtgtccg tgcagggcat
     1261 catcgtgtac cgggcctctt attttggcgc ctacgacacc gtgaagggcc tgctgcccaa
     1321 gcctaagaag acacccttcc tggtgtcttt ctttatcgcc caggtggtga ccacatgcag
     1381 cggcatcctg tcctatcctt ttgacaccgt gcggagaagg atgatgatgc agagcggcga
     1441 ggccaagaga cagtataagg gcacactgga ttgtttcgtg aagatctacc agcacgaggg
     1501 catctcctct ttctttagag gcgccttttc caatgtgctg aggggaaccg gaggcgccct
     1561 ggtgctggtg ctgtacgaca agatcaagga gttctttcac atcgatatcg gcggcaggta
     1621 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1681 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1741 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1801 aaatctctga tgttacattg cacaagataa aataatatca tcatgaacaa taaaactgtc
     1861 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1921 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1981 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2041 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2101 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2161 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2221 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2281 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2341 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2401 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2461 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2521 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2581 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2641 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2701 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2761 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2821 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2881 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2941 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3001 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3061 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3121 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3181 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3241 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3301 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3361 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3421 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3481 gccttttgct cacatgtt
//
