LOCUS       sequence-479336-        4074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4074)
  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 4074)
  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-SLC2A6_STOP
FEATURES             Location/Qualifiers
     source          1..4074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     201..220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     454..471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(630..657)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(749..835)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     884..906
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     898..915
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     899..915
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    931..1030
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2561..2660)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2677..2696)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2678..2696)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2701..2717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2701..2717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2819..2838)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2830..3639
                     /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(2897..2916)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3786..4074,1..300)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
       61 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      481 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      541 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
      601 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
      661 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
      721 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
      781 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
      841 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
      901 aaaacgacgg ccagtcttaa gctcgggccc caaataatga ttttattttg actgatagtg
      961 acctgttcgt tgcaacacat tgatgagcaa tgctttttta taatgccaac tttgtacaaa
     1021 aaagcaggct ccaccatgca ggagccactg ctgggagcag agggaccaga ctacgatacc
     1081 ttccccgaga agccacctcc aagccctggc gacagggcaa gagtgggcac cctgcagaac
     1141 aagcgggtgt tcctggccac atttgccgcc gtgctgggca atttctcttt tggctacgcc
     1201 ctggtgtata ccagccccgt gatccctgcc ctggagagat ccctggaccc tgatctgcac
     1261 ctgaccaaga gccaggcctc ctggttcggc agcgtgttca ccctgggagc agcagcagga
     1321 ggactgagcg ccatgatcct gaacgatctg ctgggcagaa agctgtccat catgttctct
     1381 gccgtgccaa gcgccgcagg atatgcactg atggcaggag cccacggcct gtggatgctg
     1441 ctgctgggaa ggaccctgac aggatttgca ggaggactga ccgcagcatg catccccgtg
     1501 tacgtgagcg agatcgcacc acctggcgtg cgcggcgccc tgggcgccac accacagctg
     1561 atggccgtgt ttggctctct gagcctgtat gccctgggcc tgctgctgcc ctggaggtgg
     1621 ctggcagtgg caggagaggc acctgtgctg atcatgatcc tgctgctgag cttcatgcca
     1681 aactccccca gatttctgct gtctagggga agggacgagg aggccctgag ggccctggcc
     1741 tggctgcgcg gcaccgacgt ggatgtgcac tgggagttcg agcagatcca ggataatgtg
     1801 cggagacaga gctccagggt gtcctgggca gaggcaaggg caccacacgt gtgcagacct
     1861 atcacagtgg ccctgctgat gcgcctgctg cagcagctga ccggcatcac acctatcctg
     1921 gtgtacctgc agtccatctt tgactctacc gccgtgctgc tgccaccaaa ggacgatgca
     1981 gcaatcgtgg gagccgtgcg gctgctgtcc gtgctgatcg ccgccctgac aatggatctg
     2041 gccggcagaa aggtgctgct gttcgtgtct gccgccatca tgtttgccgc caacctgacc
     2101 ctgggcctgt atatccactt cggccctagg ccactgtctc caaatagcac agcaggactg
     2161 gagtccgagt cttggggcga cctggcacag cctctggcag caccagcagg atacctgacc
     2221 ctggtgccac tgctggccac aatgctgttt atcatgggct atgcagtggg atggggacca
     2281 atcacctggc tgctgatgag cgaggtgctg ccactgaggg caagaggagt ggcaagcggc
     2341 ctgtgcgtgc tggcatcctg gctgaccgcc ttcgtgctga caaagtcttt tctgcctgtg
     2401 gtgagcacat tcggcctgca ggtgccattc tttttctttg ccgccatctg tctggtgagc
     2461 ctggtgttta ccggatgctg cgtgccagag acaaagggcc gctccctgga gcagatcgag
     2521 tctttcttta ggaccggcag gcgctccttc ctgcgctaag acccagcttt cttgtacaaa
     2581 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     2641 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     2701 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2761 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2821 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2881 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2941 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     3001 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     3061 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     3121 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3181 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3241 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3301 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     3361 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     3421 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     3481 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3541 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3601 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     3661 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3721 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3781 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3841 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3901 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3961 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4021 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atag
//
