LOCUS       sequence-259482-        3918 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 3918)
  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 3918)
  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_SLC29A1
FEATURES             Location/Qualifiers
     source          1..3918
                     /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(2043..2142)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2159..2178)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2160..2178)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2183..2199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2183..2199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2301..2320)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2312..3121
                     /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(2379..2398)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3268..3856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3757..3776
                     /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 accatgacca catcccacca gcctcaggac cggtacaagg ccgtgtggct
      721 gatcttcttt atgctgggcc tgggcaccct gctgccatgg aacttcttta tgaccgccac
      781 acagtatttc accaacaggc tggacatgag ccagaacgtg agcctggtga cagccgagct
      841 gagcaaggat gcacaggcat ccgccgcacc agcagcacca ctgcctgagc gcaactccct
      901 gtctgccatc tttaacaatg tgatgaccct gtgcgccatg ctgcctctgc tgctgttcac
      961 atacctgaat tcttttctgc accagcggat cccacagtct gtgagaatcc tgggcagcct
     1021 ggtggccatc ctgctggtgt tcctgatcac cgccatcctg gtgaaggtgc agctggacgc
     1081 cctgcctttc tttgtgatca caatgatcaa gatcgtgctg atcaattctt tcggcgccat
     1141 cctgcaggga agcctgtttg gactggcagg actgctgcct gcaagctaca ccgcaccaat
     1201 catgtccgga cagggactgg caggcttctt tgcaagcgtg gccatgatct gtgccatcgc
     1261 aagcggatcc gagctgtctg agagcgcctt tggctatttc atcaccgcct gcgccgtgat
     1321 catcctgaca atcatctgtt acctgggcct gcccaggctg gagttctatc gctactatca
     1381 gcagctgaag ctggagggcc ctggcgagca ggagaccaag ctggatctga tctccaaggg
     1441 agaggagcca agggcaggca aggaggagtc tggcgtgtcc gtgtctaaca gccagcccac
     1501 aaatgagtcc cactctatca aggccatcct gaagaacatc tctgtgctgg ccttcagcgt
     1561 gtgcttcatc tttaccatca caatcggcat gtttcccgcc gtgaccgtgg aggtgaagag
     1621 ctccatcgcc ggctctagca catgggagcg gtatttcatc cctgtgagct gttttctgac
     1681 cttcaatatc tttgactggc tgggcagatc cctgacagcc gtgttcatgt ggcccggcaa
     1741 ggattccagg tggctgcctt ctctggtgct ggcaaggctg gtgtttgtgc cactgctgct
     1801 gctgtgcaac atcaagcccc ggagatacct gaccgtggtg ttcgagcacg atgcctggtt
     1861 tatcttcttt atggccgcct tcgccttttc caatggctat ctggcctctc tgtgcatgtg
     1921 ctttggccca aagaaggtga agccagcaga ggcagagaca gcaggagcca tcatggcctt
     1981 cttcctgtgc ctgggactgg ccctgggagc cgtgttcagc tttctgttca gagccatcgt
     2041 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2101 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2161 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2221 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2281 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2341 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2401 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2461 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2521 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2581 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2641 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2701 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2761 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2821 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2881 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2941 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3001 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3061 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3121 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3181 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3241 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3301 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3361 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3421 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3481 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3541 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3601 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3661 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3721 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3781 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3841 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3901 gccttttgct cacatgtt
//
