LOCUS       sequence-260091-        3717 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 3717)
  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 3717)
  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_SLC14A1
FEATURES             Location/Qualifiers
     source          1..3717
                     /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(1842..1941)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1958..1977)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1959..1977)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1982..1998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..1998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2100..2119)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2111..2920
                     /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(2178..2197)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3067..3655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3556..3575
                     /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 accatggagg actcccctac catggtgagg gtggattctc caacaatggt
      721 gcgcggagag aaccaggtgt ccccatgcca gggccggaga tgttttccta aggccctggg
      781 ctatgtgacc ggcgacatga aggagctggc caatcagctg aaggacaagc ctgtggtgct
      841 gcagtttatc gattggatcc tgaggggcat ctctcaggtg gtgttcgtga acaatcccgt
      901 gagcggcatc ctgatcctgg tgggcctgct ggtgcagaac ccttggtggg cactgacagg
      961 atggctggga accgtggtga gcacactgat ggccctgctg ctgagccagg acagatccct
     1021 gatcgcctct ggcctgtatg gctacaacgc gacgctcgta ggcgtgctga tggccgtgtt
     1081 ttccgacaag ggcgattatt tctggtggct gctgctgcca gtgtgcgcca tgtctatgac
     1141 atgtcccatc ttcagctccg ccctgaactc catgctgtct aagtgggatc tgccagtgtt
     1201 caccctgccc tttaacatgg ccctgagcat gtacctgtcc gccacaggcc actataatcc
     1261 cttctttcct gccaagctgg tcatccctat caccacagcc ccaaacatca gctggtccga
     1321 cctgtctgcc ctggagctgc tgaagagcat ccctgtgggc gtgggccaga tctacggctg
     1381 cgataatcca tggaccggcg gcatctttct gggcgccatc ctgctgtcta gcccactgat
     1441 gtgcctgcac gccgccatcg gatccctgct gggaatcgca gcaggactgt ctctgagcgc
     1501 ccccttcgag gatatctact ttggcctgtg gggcttcaac tcctctctgg cctgcatcgc
     1561 catgggcggc atgttcatgg ccctgacctg gcagacacac ctgctggccc tgggctgtgc
     1621 cctgtttacc gcctatctgg gcgtgggcat ggccaatttc atggcagaag tgggactgcc
     1681 agcatgcacc tggccttttt gtctggccac actgctgttc ctgatcatga ccacaaagaa
     1741 cagcaatatc tacaagatgc cactgtccaa ggtgacatac cccgaggaga atcggatctt
     1801 ttatctgcag gccaagaaga gaatggtgga gagccccctg gacccagctt tcttgtacaa
     1861 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1921 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1981 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2041 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2101 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2161 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2221 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2281 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2341 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2401 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2461 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2521 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2581 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2641 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2701 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2761 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2821 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2881 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2941 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3001 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3061 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3121 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3601 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3661 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
