LOCUS       sequence-259271-        3597 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 3597)
  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 3597)
  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_SLC10A1
FEATURES             Location/Qualifiers
     source          1..3597
                     /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(1722..1821)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1838..1857)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1839..1857)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1862..1878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1862..1878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1980..1999)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1991..2800
                     /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(2058..2077)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2947..3535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3436..3455
                     /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 accatggagg cccacaacgc cagcgcccct tttaatttca cactgccccc
      721 taactttggc aagaggccaa ccgacctggc cctgtccgtg atcctggtgt tcatgctgtt
      781 ctttatcatg ctgagcctgg gctgcacaat ggagttttcc aagatcaagg cacacctgtg
      841 gaagccaaag ggactggcaa tcgccctggt ggcccagtac ggcatcatgc ctctgaccgc
      901 ctttgtgctg ggcaaggtgt tccggctgaa gaatatcgag gccctggcaa tcctggtgtg
      961 cggatgttct cccggcggca acctgtctaa cgtgttcagc ctggccatga agggcgacat
     1021 gaacctgtcc atcgtgatga ccacatgctc taccttttgt gccctgggca tgatgcctct
     1081 gctgctgtac atctatagca gaggcatcta cgacggcgat ctgaaggata aggtgccata
     1141 taagggcatc gtgatctccc tggtgctggt gctgatccca tgcacaatcg gcatcgtgct
     1201 gaagtctaag aggccccagt acatgcgcta tgtgatcaag ggcggcatga tcatcatcct
     1261 gctgtgcagc gtggcagtga ccgtgctgtc cgccatcaat gtgggcaagt ctatcatgtt
     1321 cgccatgaca cccctgctga tcgccaccag ctccctgatg ccttttatcg gcttcctgct
     1381 gggctatgtg ctgtccgccc tgttctgcct gaacggcaga tgtcggagaa ccgtgtctat
     1441 ggagacaggc tgccagaatg tgcagctgtg cagcacaatc ctgaacgtgg ccttcccacc
     1501 cgaagtgatc ggccctctgt tctttttccc actgctgtac atgatctttc agctgggaga
     1561 gggactgctg ctgatcgcaa tcttttggtg ttacgagaag ttcaagaccc ctaaggacaa
     1621 gacaaagatg atctataccg ccgccaccac agaggagaca atcccaggcg ccctgggcaa
     1681 tggcacctat aagggcgagg attgcagccc ctgtaccgcc gacccagctt tcttgtacaa
     1741 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1801 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1861 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1921 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1981 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2041 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2101 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2161 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2221 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2281 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2341 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2401 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2461 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2521 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2581 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2641 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2701 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2761 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2821 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2881 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2941 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3001 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3061 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3121 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3181 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3241 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3301 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3361 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3421 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3481 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3541 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
