LOCUS       sequence-263859-        4119 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 4119)
  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 4119)
  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_SLC35F5
FEATURES             Location/Qualifiers
     source          1..4119
                     /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(2244..2343)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2360..2379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2361..2379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2384..2400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2384..2400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2502..2521)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2513..3322
                     /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(2580..2599)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3469..4057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3958..3977
                     /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 accatggtgc cacctcggag acacagggga gcaggcagac ccggcgtgct
      721 gagctcctct ccacccttta gactgaggtc cgccaagttc tctggaatcg ccctggagga
      781 tctgaggcgc gccctgaaga caaggctgca gatggtgtgc gtgttcgtga tgaaccgcat
      841 gaattctcag aacagcggct ttacccagcg gagaaggatg gccctgggca tcgtgatcct
      901 gctgctggtg gacgtgatct gggtggccag ctccgagctg acctcctacg tgttcacaca
      961 gtataataag cccttctttt ccacctttgc caagacatct atgttcgtgc tgtacctgct
     1021 gggctttatc atctggaagc cttggagaca gcagtgcaca agaggcctga ggggcaagca
     1081 cgccgccttc tttgccgatg ccgagggcta cttcgccgcc tgtaccacag acaccacaat
     1141 gaactctagc ctgagcgagc ctctgtatgt gccagtgaag tttcacgatc tgcccagcga
     1201 gaagcctgag tccaccaata tcgacaccga gaagacacct aagaagtctc gcgtgcggtt
     1261 cagcaacatc atggagatcc ggcagctgcc atcctctcac gccctggagg ccaagctgtc
     1321 cagaatgtct taccccgtga aggagcagga gtctatcctg aagacagtgg gcaagctgac
     1381 cgccacacag gtggccaaga tcagcttctt tttctgtttc gtgtggttcc tggccaatct
     1441 gagctatcag gaggccctgt ccgataccca ggtggccatc gtgaacatcc tgagctccac
     1501 ctccggcctg tttacactga tcctggccgc cgtgttccca tccaattctg gcgacaggtt
     1561 tacactgtcc aagctgctgg ccgtgatcct gtctatcgga ggagtggtgc tggtgaacct
     1621 ggcaggaagc gagaagccag caggacgcga taccgtggga agcatctggt ccctggcagg
     1681 agcaatgctg tacgccgtgt atatcgtgat gatcaagcgg aaggtggaca gagaggataa
     1741 gctggacatc cctatgtttt tcggctttgt gggcctgttc aatctgctgc tgctgtggcc
     1801 aggctttttc ctgctgcact acacaggctt cgaggacttt gagttcccca ataaggtggt
     1861 gctgatgtgc atcatcatca acggcctgat cggcaccgtg ctgagcgagt ttctgtggct
     1921 gtggggctgt ttcctgacct ctagcctgat cggcacactg gccctgtctc tgaccatccc
     1981 tctgagcatc atcgccgata tgtgcatgca gaaggtgcag ttcagctggc tgtttttcgc
     2041 cggcgccatc cccgtgttct tcagcttctt catcgtgacc ctgctgtgcc actataacaa
     2101 ttgggaccca gtgatggtgg gcatccgccg gatctttgcc ttcatctgcc gcaagcaccg
     2161 gatccagaga gtgccagagg attccgagca gtgtgagtct ctgatcagca tgcacagcgt
     2221 gtcccaggag gacggagcat ctgacccagc tttcttgtac aaagttggca ttataagaaa
     2281 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2341 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2401 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2461 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2521 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2581 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2641 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2701 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2761 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2821 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2881 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2941 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3001 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3061 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3121 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3181 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3241 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3301 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3361 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3421 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3481 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3541 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4081 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
