LOCUS       sequence-263855-        3855 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 3855)
  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 3855)
  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_SLC38A8
FEATURES             Location/Qualifiers
     source          1..3855
                     /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(1980..2079)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2096..2115)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2097..2115)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2120..2136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2120..2136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2238..2257)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2249..3058
                     /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(2316..2335)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3205..3793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3694..3713
                     /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 gacagacccc aggatccagg ggactgcctg agaagccaca
      721 ccccgccacc gccgccgcca cactgagctc catgggcgcc gtgttcatcc tgatgaagtc
      781 cgccctggga gcaggactgc tgaacttccc ttgggccttt tctaaggcag gaggagtggt
      841 gccagccttc ctggtggagc tggtgtccct ggtgtttctg atctctggac tggtcatcct
      901 gggatacgca gcagccgtgt ctggacaggc cacatatcag ggagtggtgc ggggactgtg
      961 cggaccagcc atcggcaagc tgtgcgaggc ctgtttcctg ctgaatctgc tgatgatcag
     1021 cgtggccttt ctgagagtga tcggcgacca gctggagaag ctgtgcgatt ctctgctgag
     1081 cggaacccca cctgcacctc agccatggta cgcagaccag cggttcacac tgcctctgct
     1141 gagcgtgctg gtcatcctgc cactgtctgc ccccagagag atcgcctttc agaagtacac
     1201 cagcatcctg ggcacactgg cagcatgcta cctggccctg gtcatcaccg tgcagtacta
     1261 tctgtggcct cagggcctgg tgagagagtc ccacccttcc ctgtctccag cctcctggac
     1321 cagcgtgttc agcgtgttcc caacaatctg cttcggcttt cagtgtcacg aggccgccgt
     1381 gagcatctat tgttccatga ggaagcgcag cctgtcccac tgggcactgg tgtccgtgct
     1441 gtctctgctg gcctgctgtc tgatctactc cctgaccggc gtgtatggct tcctgacctt
     1501 tggcacagag gtgagcgccg acgtgctgat gtcctaccca ggcaacgata tggtcatcat
     1561 cgtggccagg gtgctgttcg ccgtgagcat cgtgacagtg tatcccatcg tgctgttcct
     1621 gggccgcagc gtgatgcagg acttttggcg gagatcctgc ctgggaggat ggggaccatc
     1681 tgccctggca gatccaagcg gactgtgggt gaggatgcca ctgaccatcc tgtgggtgac
     1741 cgtgacactg gccatggccc tgtttatgcc tgatctgtct gagatcgtga gcatcatcgg
     1801 cggcatctct agcttcttta tcttcatctt tcccggcctg tgcctgatct gtgcaatggg
     1861 agtggagcca atcggaccta gggtgaagtg ctgtctggaa gtgtggggcg tggtgtctgt
     1921 gctggtgggc accttcatct ttggacagag cacagcagca gccgtgtggg agatgttcga
     1981 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2041 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2101 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2161 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2221 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2281 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2341 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2401 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2461 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2521 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2581 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2641 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2701 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2761 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2821 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2881 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2941 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3001 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3061 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3121 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3841 ttttgctcac atgtt
//
