LOCUS       sequence-262961-        3663 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 3663)
  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 3663)
  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_SLC35F6
FEATURES             Location/Qualifiers
     source          1..3663
                     /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(1788..1887)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1904..1923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1905..1923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1928..1944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1928..1944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2046..2065)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2057..2866
                     /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(2124..2143)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3013..3601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3502..3521
                     /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 accatggcct ggaccaagta ccagctgttt ctggcaggac tgatgctggt
      721 gaccggaagc atcaacacac tgtccgccaa gtgggctgat aatttcatgg cggaaggctg
      781 cggaggatct aaggagcaca gcttccagca cccttttctg caggccgtgg gcatgttcct
      841 gggcgagttt agctgcctgg ccgcctttta cctgctgagg tgtagggcag caggacagtc
      901 cgacagctcc gtggatccac agcagccctt taaccctctg ctgttcctgc cacccgccct
      961 gtgcgatatg accggcacaa gcctgatgta tgtggccctg aatatgacct ccgcctctag
     1021 cttccagatg ctgcggggcg ccgtgatcat cttcacaggc ctgttttccg tggccttcct
     1081 gggccggaga ctggtgctgt ctcagtggct gggaatcctg gcaaccatcg caggactggt
     1141 ggtggtggga ctggcagacc tgctgtccaa gcacgattct cagcacaagc tgtctgaagt
     1201 gatcacaggc gacctgctga tcatcatggc ccagatcatc gtggccatcc agatggtgct
     1261 ggaggagaag ttcgtgtaca agcacaacgt gcacccactg agggcagtgg gaaccgaggg
     1321 cctgttcggc tttgtgatcc tgtccctgct gctggtgcca atgtactata tccccgccgg
     1381 cagcttttcc ggcaatccca gaggcacact ggaggacgcc ctggatgcct tctgccaagt
     1441 gggacagcag cctctgatcg cagtggccct gctgggcaac atctcctcta tcgccttctt
     1501 taatttcgcc ggcatctctg tgaccaagga gctgagcgcc accacaagga tggtgctgga
     1561 ctctctgcgc acagtggtca tctgggcact gagcctggcc ctgggatggg aggcatttca
     1621 cgccctgcag atcctgggct tcctgatcct gctgatcggc accgccctgt ataacggcct
     1681 gcacaggcct ctgctgggcc gcctgtccag gggcagacca ctggcagagg agtctgagca
     1741 ggagcggctg ctgggaggaa ccagaacacc catcaatgat gccagcgacc cagctttctt
     1801 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1861 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     1921 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1981 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2041 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2101 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2161 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2221 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2281 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2341 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2401 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2461 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2521 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2581 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2641 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2701 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2761 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2821 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2881 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     2941 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3001 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3061 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3121 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3181 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3241 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3301 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3361 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3421 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3481 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3541 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3601 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3661 gtt
//
