LOCUS       sequence-265082-        4113 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 4113)
  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 4113)
  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_SLC35F4
FEATURES             Location/Qualifiers
     source          1..4113
                     /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(2238..2337)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2354..2373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2355..2373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2378..2394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2378..2394)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2496..2515)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2507..3316
                     /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(2574..2593)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3463..4051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3952..3971
                     /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 accatggacg agctgctgct ggatctgttc cacaagctga ccagcggcag
      721 gcagctggca gcaggaaacg gactgtgcgg catctcccac aaggagcagg aagtgtggaa
      781 gcctggccac aatatcctgg tgaagatgag gaaggaggac aagtccctgg tgtggctgat
      841 ccactctacc ctggcccgct acacccaggt gacaaacttt ctgggcacct ccaggagctc
      901 cgtgacaagg tgcaagccag gagccaattg tccttctagc cacagcggca tctcccgcca
      961 gctgtctcca ctgagcgtga cagaggattc ctctgccccc atcctggagc tgcagaacca
     1021 gggcagctcc ggcgtgtgcg gacacagggt ggagaggcag aatagaagcg ccgacgatgg
     1081 cacccagaca cactccgaga actctagcca ggagaatagg atcaaggccc gctgcctgtc
     1141 ctgtacctct atggtgctga agggcatctg gggcctgctg atcatcctga gcgtgtcctc
     1201 tagctgggtg ggcaccacac agatcgtgaa gatcacatac aagaacttct attgcccctt
     1261 ctttatgacc tggttttcca caaactggaa tatcatgttc tttcccgtgt actattctgg
     1321 acacctggca accgcacagg agaagcagtc tccaatgaag aagttccggg agtgtagcag
     1381 aatctttggc gaggacggcc tgaccctgaa gctgttcctg aagcggacag ccccattttc
     1441 catcctgtgg accctgacaa actacctgta tctgctggcc ctgaagaagc tgaccgccac
     1501 agacgtgtct gccctgttct gctgtaataa ggccttcgtg tttctgctga gctggatcgt
     1561 gctgaaggat cggtttatgg gcgtgagaat cgtggccgcc atcatggcca tcaccggcat
     1621 cgtgatgatg gcctacgccg acaatttcca cgccgatagc atcatcggag tggcctttgc
     1681 cgtgggaagc gcctccacct ctgccctgta taaggtgctg ttcaagatgt ttctgggatc
     1741 cgccaacttc ggagaggcag cacacttcgt gagcaccctg ggcttcttta atctgatctt
     1801 catcagcttt accccagtga tcctgtactt cacaaaggtg gagcactggt cctcttttgc
     1861 cgccctgcca tggggatgcc tgtgcggaat ggccggcctg tggctggcct tcaacatcct
     1921 ggtgaatgtg ggcgtggtgc tgacctatcc tatcctgatc agcatcggca cagtgctgtc
     1981 cgtgccagga aacgcagcag tggatctgct gaagcaggaa gtgatcttca acgtggtgcg
     2041 gctggccgcc accatcatca tctgcatcgg ctttctgctg atgctgctgc ctgaggagtg
     2101 ggacgagatc acactgagat tcatcaactc tctgaaggag aagaagagcg aggagcacgt
     2161 ggacgatgtg accgatccat ccatccacct gaggggaaga ggaagggcaa atggaacagt
     2221 gagcatccct ctggccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2281 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2341 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2401 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2461 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2521 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2581 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2641 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2701 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2761 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2821 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2881 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2941 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3001 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3061 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3121 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3181 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3241 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3301 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3361 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3601 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4081 cctggccttt tgctggcctt ttgctcacat gtt
//
