LOCUS       sequence-261923-        4575 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 4575)
  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 4575)
  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_SLC5A11
FEATURES             Location/Qualifiers
     source          1..4575
                     /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(2700..2799)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2816..2835)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2817..2835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2840..2856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2840..2856)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2958..2977)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2969..3778
                     /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(3036..3055)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3925..4513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4414..4433
                     /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 accatggagt ccggcaccag ctcccctcag ccacctcagc tggacccact
      721 ggatgcattc cctcagaagg gcctggagcc aggcgatatc gccgtgctgg tgctgtactt
      781 cctgtttgtg ctggcagtgg gactgtggag caccgtgaag acaaagcggg acaccgtgaa
      841 gggctatttt ctggcaggag gcgatatggt gtggtggcca gtgggagcat ctctgttcgc
      901 cagcaacgtg ggatccggac actttatcgg actggcagga tctggagcag caacaggaat
      961 ctccgtgtct gcctacgagc tgaatggcct gttcagcgtg ctgatgctgg cctggatctt
     1021 tctgccaatt tatatcgccg gccaggtgac cacaatgccc gagtatctgc ggaagagatt
     1081 cggcggcatc aggatcccta tcatcctggc cgtgctgtac ctgttcatct atatctttac
     1141 caagatcagc gtggacatgt acgccggcgc catctttatc cagcagtccc tgcacctgga
     1201 tctgtatctg gcaatcgtgg gactgctggc aatcaccgcc gtgtacacag tggcaggagg
     1261 actggcagcc gtgatctata ccgacgccct gcagacactg atcatgctga tcggcgccct
     1321 gaccctgatg ggatactcct tcgccgcagt gggaggaatg gagggcctga aggagaagta
     1381 ttttctggcc ctggcctcta accggagcga gaattctagc tgcggactgc ctagggagga
     1441 cgcattccac atctttcgcg acccactgac aagcgatctg ccatggcccg gcgtgctgtt
     1501 cggcatgtcc atcccttctc tgtggtactg gtgtaccgat caggtcatcg tgcagaggac
     1561 actggccgcc aagaacctgt ctcacgcaaa gggaggcgcc ctgatggcag catatctgaa
     1621 ggtgctgcca ctgttcatca tggtgtttcc cggcatggtg agccgcatcc tgtttcctga
     1681 ccaggtggcc tgcgccgatc cagagatctg ccagaagatc tgtagcaatc catccggctg
     1741 ttctgacatc gcctacccca agctggtgct ggagctgctg cctaccggac tgaggggact
     1801 gatgatggcc gtgatggtgg ccgccctgat gtcctctctg acaagcatct tcaacagcgc
     1861 ctccaccatc tttacaatgg atctgtggaa tcacctgagg ccaagggcat ccgagaagga
     1921 gctgatgatc gtgggcagag tgttcgtgct gctgctggtg ctggtgtcca tcctgtggat
     1981 ccctgtggtg caggcatctc agggaggaca gctgtttatc tacatccagt ccatcagctc
     2041 ctatctgcag ccacccgtgg ccgtggtgtt catcatgggc tgcttttgga agcggaccaa
     2101 cgagaaggga gcattctggg gactgatctc cggactgctg ctgggactgg tgagactggt
     2161 gctggacttc atctacgtgc agccccggtg tgaccagcct gatgagagac cagtgctggt
     2221 gaagtctatc cactacctgt atttctctat gatcctgagc accgtgacac tgatcaccgt
     2281 gagcacagtg tcctggttca cagagcctcc atctaaggag atggtgagcc acctgacctg
     2341 gtttacaagg cacgacccag tggtgcagaa ggagcaggca ccacctgcag caccactgtc
     2401 cctgaccctg tctcagaacg gcatgcctga ggcctctagc tcctctagcg tgcagttcga
     2461 gatggtgcag gagaatacca gcaagacaca ctcctgcgat atgacaccca agcagagcaa
     2521 ggtggtgaag gccatcctgt ggctgtgcgg catccaggag aagggcaagg aggagctgcc
     2581 agcaagggca gaggcaatca tcgtgtccct ggaggagaac cccctggtga agaccctgct
     2641 ggacgtgaat ctgatcttct gcgtgtcttg tgccatcttc atctggggct actttgccga
     2701 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2761 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2821 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2881 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2941 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     3001 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     3061 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     3121 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     3181 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     3241 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     3301 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3361 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3421 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3481 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3541 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3601 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3661 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3721 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3781 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3841 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3901 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3961 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4021 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4081 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4141 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4201 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4261 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4321 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4381 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4441 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4501 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4561 ttttgctcac atgtt
//
