LOCUS       sequence-260220-        3546 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 3546)
  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 3546)
  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_SLC25A16
FEATURES             Location/Qualifiers
     source          1..3546
                     /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(1671..1770)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1787..1806)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1788..1806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1811..1827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1811..1827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1929..1948)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1940..2749
                     /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(2007..2026)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2896..3484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3385..3404
                     /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 accatggcag cagcaaccgc agcagccgcc ctggccgccg ccgacccccc
      721 tccagccatg cctcaggctg ctggcgccgg cggcccaacc acacggagag atttctactg
      781 gctgagatct tttctggcag gaggaatcgc aggatgctgt gcaaagacca cagtggcccc
      841 cctggacagg gtgaaggtgc tgctgcaggc ccacaaccac cactacaagc acctgggcgt
      901 gttcagcgcc ctgagggcag tgccacagaa ggagggcttt ctgggcctgt ataagggcaa
      961 tggcgccatg atgatccgga tcttccccta cggcgccatc cagttcatgg cctttgagca
     1021 ctataagaca ctgatcacca caaagctggg catcagcgga cacgtgcaca ggctgatggc
     1081 aggatccatg gcaggaatga ccgccgtgat ctgcacatac cctctggaca tggtgagagt
     1141 gaggctggcc ttccaggtga agggcgagca ctcctatacc ggcatcatcc acgccttcaa
     1201 gacaatctac gccaaggagg gcggcttctt tggcttttat cgcggcctga tgccaaccat
     1261 cctgggaatg gcaccatacg caggcgtgtc tttctttacc tttggcacac tgaagtctgt
     1321 gggactgagc cacgcaccaa cactgctggg cagacccagc tccgataacc ctaatgtgct
     1381 ggtgctgaag acccacgtga acctgctgtg cggaggagtg gcaggagcaa tcgcccagac
     1441 catcagctat cccttcgacg tgacaaggcg ccggatgcag ctgggaaccg tgctgcctga
     1501 gtttgagaag tgcctgacca tgagagatac aatgaagtac gtgtatggcc accacggaat
     1561 caggaaggga ctgtacaggg gactgtccct gaactatatc aggtgtatcc cttctcaggc
     1621 cgtggccttt accacatacg agctgatgaa gcagttcttt cacctgaatg acccagcttt
     1681 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     1741 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     1801 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     1861 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1921 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1981 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2041 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2101 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2161 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2221 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2281 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2341 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2401 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2461 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2521 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2581 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2641 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2701 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2761 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     2821 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     2881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3541 catgtt
//
