LOCUS       sequence-260102-        4317 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 4317)
  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 4317)
  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_SLC17A8
FEATURES             Location/Qualifiers
     source          1..4317
                     /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(2442..2541)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2558..2577)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2559..2577)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2582..2598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2582..2598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2700..2719)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2711..3520
                     /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(2778..2797)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3667..4255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4156..4175
                     /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 accatgccat tcaaggcctt tgacacattc aaggagaaga tcctgaagcc
      721 cggcaaggag ggcgtgaaga acgccgtggg cgatagcctg ggcatcctgc agaggaagat
      781 cgacggcacc acagaggagg aggataacat cgagctgaat gaggagggca ggcccgtgca
      841 gacctccagg ccttctcccc ctctgtgcga ctgtcactgc tgtggcctgc ctaagaggta
      901 catcatcgcc atcatgtctg gcctgggctt ttgcatcagc ttcggcatcc gctgtaacct
      961 gggcgtggcc atcgtggaga tggtgaacaa ttccacagtg tacgtggacg gcaagcccga
     1021 gatccagaca gcccagttta attgggatcc tgagaccgtg ggcctgatcc acggcagctt
     1081 cttttggggc tatatcatga cccagatccc aggcggcttt atctccaaca agttcgccgc
     1141 caatagagtg tttggcgccg ccatcttcct gacatccacc ctgaatatgt tcatcccatc
     1201 tgccgcaagg gtgcactacg gatgcgtgat gtgcgtgcgg atcctgcagg gactggtgga
     1261 gggcgtgacc tatcctgcct gccacggcat gtggtctaag tgggcaccac cactggagcg
     1321 gagcagactg gccaccacat ccttttgtgg atcttacgca ggagcagtgg tggcaatgcc
     1381 actggccggc gtgctggtgc agtatatcgg ctggagctcc gtgttttaca tctatggcat
     1441 gttcggcatc atctggtaca tgttctggct gctgcaggca tatgagtgcc ctgcagcaca
     1501 cccaaccatc tctaacgagg agaagacata catcgagacc agcatcggcg agggcgccaa
     1561 tgtggtgtct ctgagcaagt tttccacacc atggaagagg ttctttacct ctctgcccgt
     1621 gtatgccatc atcgtggcca acttttgtcg cagctggaca ttctacctgc tgctgatctc
     1681 ccagcctgcc tatttcgagg aggtgttcgg ctttgccatc tccaaagtgg gcctgctgtc
     1741 tgccgtgcct cacatggtca tgaccatcgt ggtgccaatc ggaggacagc tggcagacta
     1801 cctgaggagc cgccagatcc tgaccacaac cgccgtgcgg aagatcatga attgcggcgg
     1861 ctttggcatg gaggccacac tgctgctggt ggtgggcttt agccacacca agggcgtggc
     1921 catctccttc ctggtgctgg ccgtgggctt tagcggcttc gccatctccg gcttcaacgt
     1981 gaatcacctg gatatcgccc ctagatatgc cagcatcctg atgggcatct ccaacggcgt
     2041 gggcaccctg tctggcatgg tgtgcccact gatcgtgggc gccatgacac ggcacaagac
     2101 cagagaggag tggcagaacg tgttcctgat cgccgccctg gtgcactact ccggcgtgat
     2161 cttttatggc gtgttcgcct ctggcgagaa gcaggagtgg gccgacccag agaacctgtc
     2221 cgaggagaag tgcggcatca tcgaccagga tgagctggcc gaggagatcg agctgaacca
     2281 cgagagcttt gcctccccca agaagaagat gtcttacggc gccacaagcc agaattgtga
     2341 ggtgcagaag aaggagtgga agggacagag gggagccaca ctggatgagg aggagctgac
     2401 cagctatcag aacgaggaga gaaatttctc taccatcagc gacccagctt tcttgtacaa
     2461 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2521 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2581 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2641 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2701 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2761 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2821 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2881 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2941 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3001 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3061 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3121 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3181 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3241 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3301 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3361 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3421 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3481 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3541 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3601 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3781 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4021 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
