LOCUS       sequence-260202-        3804 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 3804)
  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 3804)
  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_SLC25A46
FEATURES             Location/Qualifiers
     source          1..3804
                     /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(1929..2028)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2045..2064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2046..2064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2069..2085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2069..2085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2187..2206)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2198..3007
                     /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(2265..2284)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3643..3662
                     /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 cccggagacc tgacggcttc gatggactgg gatacagggg
      721 aggagcaaga gacgagcagg gattcggagg agcatttcca gcacggagct tttccaccgg
      781 aagcgacctg ggacactggg tgaccacacc acctgatatc cccggctcca gaaacctgca
      841 ctggggcgag aagtctccac cctacggcgt gcctaccaca agcaccccat atgagggccc
      901 cacagaggag cctttcagct ccggaggagg aggatctgtg cagggacagt ctagcgagca
      961 gctgaatagg ttcgcaggat ttggaatcgg actggcatcc ctgtttacag agaacgtgct
     1021 ggcccaccct tgcatcgtgc tgaggcgcca gtgtcaagtg aattaccacg cccagcacta
     1081 tcacctgacc cctttcacag tgatcaacat catgtactct tttaataaga cccagggacc
     1141 aagggccctg tggaagggaa tgggcagcac cttcatcgtg cagggagtga cactgggagc
     1201 agagggcatc atctccgagt ttacacctct gccacgcgag gtgctgcaca agtggagccc
     1261 aaagcagatc ggcgagcacc tgctgctgaa gtccctgacc tacgtggtgg ccatgccctt
     1321 ctattctgcc agcctgatcg agaccgtgca gagcgagatc atccgggaca atacaggcat
     1381 cctggagtgc gtgaaggagg gcatcggcag agtgatcggc atgggcgtgc cacactctaa
     1441 gcgcctgctg cccctgctga gcctgatctt ccctaccgtg ctgcacggcg tgctgcacta
     1501 catcatctcc tctgtgatcc agaagtttgt gctgctgatc ctgaagagga agacctacaa
     1561 ctcccacctg gccgagtcca catctccagt gcagtctatg ctggacgcct atttccccga
     1621 gctgatcgcc aattttgccg ccagcctgtg ctccgatgtg atcctgtatc ctctggagac
     1681 agtgctgcac aggctgcaca tccagggcac ccgcacaatc atcgacaaca ccgatctggg
     1741 ctacgaggtg ctgccaatca atacacagta tgagggcatg cgggattgta tcaacaccat
     1801 cagacaggag gagggcgtgt tcggctttta caagggcttt ggcgccgtga tcatccagta
     1861 taccctgcac gccgccgtgc tgcagatcac caagatcatc tactccacac tgctgcagaa
     1921 caatatcgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1981 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2041 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2101 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2161 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2221 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2281 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2341 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2401 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2461 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2521 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2581 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2641 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2701 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2761 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2821 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2881 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2941 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3001 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3061 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgtt
//
