LOCUS       sequence-262930-        3489 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 3489)
  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 3489)
  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_SLC35E3
FEATURES             Location/Qualifiers
     source          1..3489
                     /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(1614..1713)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1730..1749)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1731..1749)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1754..1770)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1754..1770)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1872..1891)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1883..2692
                     /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(1950..1969)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2839..3427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3328..3347
                     /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 accatggcac tgctggtgga ccgggtgaga ggacactgga ggatcgcagc
      721 aggactgctg tttaacctgc tggtgtccat ctgcatcgtg ttcctgaata agtggatcta
      781 cgtgtatcac ggctttccta acatgtctct gaccctggtg cacttcgtgg tgacatggct
      841 gggcctgtac atctgccaga agctggatat ctttgcacca aagagcctgc caccttccag
      901 gctgctgctg ctggccctgt ccttctgtgg ctttgtggtg ttcaccaatc tgtctctgca
      961 gaacaatacc atcggcacat accagctggc caaggccatg accacacctg tgatcatcgc
     1021 catccagaca ttttgttatc agaagacctt ctctacaagg atccagctga ccctgatccc
     1081 aatcacactg ggcgtgatcc tgaacagcta ctatgacgtg aagttcaatt ttctgggaat
     1141 ggtgttcgcc gccctgggcg tgctggtgac cagcctgtat caagtgtggg tgggagcaaa
     1201 gcagcacgag ctgcaggtga actccatgca gctgctgtac tatcaggccc ccatgagctc
     1261 cgccatgctg ctggtggcag tgccattctt tgagcccgtg tttggcgagg gcggcatctt
     1321 cggcccttgg tctgtgagcg ccctgctgat ggtgctgctg agcggcgtga tcgcctttat
     1381 ggtgaatctg tctatctact ggatcatcgg caacaccagc ccagtgacat ataatatgtt
     1441 cggccacttc aagttttgca tcaccctgtt tggcggctac gtgctgttca aggatcccct
     1501 gagcatcaac caggccctgg gcatcctgtg caccctgttt ggcatcctgg cctatacaca
     1561 cttcaagctg tccgagcagg agggctccag gtctaagctg gcccagcgcc ccgacccagc
     1621 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1681 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1741 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1801 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1861 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1921 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1981 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2041 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2101 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2161 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2221 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2281 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2341 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2401 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2461 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2521 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2581 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2641 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2701 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2761 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3481 tcacatgtt
//
