LOCUS       sequence-263891-        3822 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 3822)
  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 3822)
  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_SLC35A5
FEATURES             Location/Qualifiers
     source          1..3822
                     /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(1947..2046)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2063..2082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2064..2082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2087..2103)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2087..2103)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2205..2224)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2216..3025
                     /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(2283..2302)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3172..3760
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3661..3680
                     /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 accatggaga agcagtgctg tagccaccct gtgatctgct ccctgtctac
      721 catgtacaca ttcctgctgg gcgccatctt tatcgccctg agctcctctc ggatcctgct
      781 ggtcaagtat agcgccaacg aggagaataa gtacgactat ctgccaacca cagtgaacgt
      841 gtgctccgag ctggtgaagc tggtgttctg cgtgctggtg tccttttgcg tgatcaagaa
      901 ggatcaccag tctaggaatc tgaagtacgc ctcttggaag gagttcagcg actttatgaa
      961 gtggtctatc cccgccttcc tgtattttct ggataacctg atcgtgttct acgtgctgag
     1021 ctatctgcag cctgccatgg ccgtgatctt cagcaatttt tccatcatca ccacagccct
     1081 gctgttcagg atcgtgctga agcggagact gaactggatc cagtgggcca gcctgctgac
     1141 cctgtttctg tccatcgtgg ccctgacagc cggcaccaag acactgcagc acaacctggc
     1201 aggaagggga ttccaccacg acgccttctt tagcccttcc aactcttgcc tgctgtttag
     1261 gagcgagtgc ccacgcaagg ataattgtac cgccaaggag tggacattcc ccgaggccaa
     1321 gtggaacacc acagcccggg tgttttccca catcagactg ggcatgggcc acgtgctgat
     1381 catcgtgcag tgtttcatca gctccatggc caacatctac aatgagaaga tcctgaagga
     1441 gggcaaccag ctgaccgagt ccatcttcat ccagaattct aagctgtatt tctttggcat
     1501 cctgtttaat ggcctgacac tgggcctgca gcggagcaac agagaccaga tcaagaattg
     1561 cggcttcttt tacggccact ctgccttcag cgtggccctg atcttcgtga ccgcctttca
     1621 gggcctgtcc gtggccttca tcctgaagtt tctggacaac atgtttcacg tgctgatggc
     1681 ccaggtgacc acagtgatca tcaccacagt gtctgtgctg gtgttcgatt ttcggccaag
     1741 cctggagttc tttctggagg cccccagcgt gctgctgtcc atcttcatct acaatgcctc
     1801 caagccccag gtgcctgagt atgccccaag acaggagagg atccgcgacc tgtctggcaa
     1861 cctgtgggag aggtctagcg gcgatggaga ggagctggag cgcctgacca agccaaagtc
     1921 cgatgagtct gacgaggata catttgaccc agctttcttg tacaaagttg gcattataag
     1981 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2041 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2101 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2161 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2221 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2281 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2341 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2401 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2461 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2521 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2581 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2641 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2701 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2761 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2821 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2881 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2941 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3001 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3061 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3121 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3181 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3241 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3301 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3361 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3421 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3481 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3541 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3601 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3661 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3721 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3781 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
