LOCUS       sequence-259278-        3810 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 3810)
  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 3810)
  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_SLC17A3
FEATURES             Location/Qualifiers
     source          1..3810
                     /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"
     regulatory      858..867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1935..2034)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2051..2070)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2052..2070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2075..2091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2075..2091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2193..2212)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2204..3013
                     /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(2271..2290)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3160..3748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3649..3668
                     /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 accatggcca ccaagacaga gctgtcccct accgcccggg agtctaagaa
      721 cgcccaggac atgcaggtgg atgagacact gatccctagg aaggtgccat ctctgtgcag
      781 cgccagatac ggaatcgccc tggtgctgca cttctgtaac ttcaccacaa tcgcccagaa
      841 cgtgatcatg aatatcacca tggtggccat ggtgaactcc acatctccac agagccagct
      901 gaatgacagc tccgaggtgc tgcccgtgga tagcttcggc ggcctgtcca aggcccccaa
      961 gtctctgcct gccaagtcta gcatcctggg cggccagttt gccatctggg agaagtgggg
     1021 accacctcag gagcggtcca gactgtgctc tatcgccctg agcggcatgc tgctgggctg
     1081 tttcaccgcc atcctgatcg gcggctttat cagcgagaca ctgggctggc cattcgtgtt
     1141 ttacatcttc ggaggagtgg gatgcgtgtg ctgtctgctg tggtttgtgg tcatctacga
     1201 cgatccagtg tcctatccct ggatcagcac ctccgagaag gagtatatca tctcctctct
     1261 gaagcagcaa gtgggcagct ccaagcagcc actgcccatc aaggccatgc tgaggagcct
     1321 gcccatctgg tccatctgcc tgggctgttt cagccaccag tggctggtgt ccaccatggt
     1381 ggtgtacatc cctacatata tctctagcgt gtaccacgtg aacatcaggg acaatggact
     1441 gctgagcgcc ctgcccttca tcgtggcatg ggtcatcgga atggtgggag gctatctggc
     1501 cgatttcctg ctgaccaaga agtttcggct gatcaccgtg agaaagatcg ccacaatcct
     1561 gggctccctg ccctcctctg ccctgatcgt gtctctgcct tacctgaaca gcggctatat
     1621 caccgcaaca gccctgctga ccctgtcctg cggactgagc accctgtgcc agtccggcat
     1681 ctacatcaat gtgctggaca tcgccccccg gtatagctcc ttcctgatgg gcgcctctag
     1741 aggcttttct agcatcgccc ctgtgatcgt gccaaccgtg tctggcttcc tgctgagcca
     1801 ggaccctgag tttggctgga gaaacgtgtt ctttctgctg ttcgccgtga atctgctggg
     1861 cctgctgttc tatctgatct ttggcgaggc cgatgtgcag gagtgggcca aggagaggaa
     1921 gctgacacgc ctggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2041 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2101 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2161 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2221 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2281 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2341 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2401 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2461 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2521 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2581 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2641 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2701 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2761 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2821 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2881 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2941 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3001 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3061 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3781 ggccttttgc tggccttttg ctcacatgtt
//
