LOCUS       sequence-259493-        3981 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 3981)
  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 3981)
  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_SLC10A3
FEATURES             Location/Qualifiers
     source          1..3981
                     /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(2106..2205)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2222..2241)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2223..2241)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2246..2262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2246..2262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2364..2383)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2375..3184
                     /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(2442..2461)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3331..3919
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3820..3839
                     /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 accatggtgc tgatgcagga caagggcagc tcccagcagt ggcctggact
      721 gggaggagag ggaggaggaa ccggaccact gtccatgctg agggccgccc tgctgctgat
      781 ctctctgcca tggggagcac agggaaccgc atccacatct ctgagcacag caggaggaca
      841 caccgtgcca cctacaggag gccgctacct gagcatcggc gacggatccg tgatggagtt
      901 cgagtttcca gaggattccg agggcatcat cgtgatctct agccagtatc caggacaggc
      961 aaacaggacc gcacctggac caatgctgag ggtgacctct ctggatacag aggtgctgac
     1021 catcaagaat gtgagcgcca tcacatgggg cggaggagga ggcttcgtgg tgtccatcca
     1081 ctctggactg gcaggactgg caccactgca catccagctg gtggacgcac acgaggcacc
     1141 acccaccctg atcgaggaga ggagagattt ctgcatcaag gtgagccctg cagaggacac
     1201 cccagccaca ctgagcgccg atctggcaca cttttccgag aaccccatcc tgtacctgct
     1261 gctgcctctg atcttcgtga ataagtgctc ctttggctgt aaggtggagc tggaggtgct
     1321 gaagggcctg atgcagtctc cacagcctat gctgctggga ctgctgggac agttcctggt
     1381 catgcccctg tacgccttcc tgatggccaa ggtgtttatg ctgcctaagg ccctggccct
     1441 gggcctgatc atcacctgtt cctctcctgg aggaggagga tcttatctgt ttagcctgct
     1501 gctgggaggc gacgtgacac tggccatctc tatgacattc ctgagcaccg tggccgccac
     1561 aggctttctg cctctgagct ccgccatcta ttcccggctg ctgtctatcc acgagaccct
     1621 gcacgtgcca atctccaaga tcctgggcac actgctgttc atcgccatcc ccatcgccgt
     1681 gggcgtgctg atcaagtcca agctgcctaa gttttctcag ctgctgctgc aggtggtgaa
     1741 gccattcagc tttgtgctgc tgctgggagg actgttcctg gcatacagga tgggcgtgtt
     1801 tatcctggcc ggcatcagac tgccaatcgt gctggtggga atcaccgtgc cactggtggg
     1861 actgctggtg ggctattgcc tggccacctg tctgaagctg ccagtggcac agaggaggac
     1921 agtgtctatc gaagtgggag tgcagaacag cctgctggcc ctggcaatgc tgcagctgag
     1981 cctgcggaga ctgcaggccg attacgcctc ccaggcccct ttcatcgtgg ccctgagcgg
     2041 cacctccgag atgctggccc tggtcatcgg ccacttcatc tattctagcc tgtttccagt
     2101 gcccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2161 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2221 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2281 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2341 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2401 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2461 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2521 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2581 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2641 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2701 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2761 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2821 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2881 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2941 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3001 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3061 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3121 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3181 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3241 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3301 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3361 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3421 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3481 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3541 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3601 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3661 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3721 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3781 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3841 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3901 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3961 ctggcctttt gctcacatgt t
//
