LOCUS       sequence-259459-        4065 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 4065)
  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 4065)
  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_SLC16A5
FEATURES             Location/Qualifiers
     source          1..4065
                     /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(2190..2289)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2306..2325)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2307..2325)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2330..2346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2330..2346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2448..2467)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2459..3268
                     /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(2526..2545)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3415..4003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3904..3923
                     /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 accatgccac aggccctgga gagggcagac ggatcctggg catgggtggt
      721 gctgctggca accatggtga cacagggcct gaccctgggc ttccccacat gcatcggcat
      781 cttctttacc gagctgcagt gggagttcca ggcctccaac tctgagacca gctggtttcc
      841 ttccatcctg acagccgtgc tgcacatggc aggaccactg tgctccatcc tggtgggcag
      901 attcggctgt agggtgacag tgatgctggg aggcgtgctg gcatctctgg gcatggtggc
      961 cagctccttt tctcacaatc tgagccagct gtacttcacc gccggcttta tcacaggcct
     1021 gggcatgtgc ttcagctttc agtctagcat caccgtgctg ggcttctatt ttgtgcggag
     1081 aagggtgctg gcaaacgcac tggcatccat gggcgtgtct ctgggcatca ccctgtggcc
     1141 tctgctgtct cgctacctgc tggagaatct gggctggcgg ggcacattcc tggtgtttgg
     1201 cggcatcttc ctgcactgct gtatctgcgg cgccatcatc aggccagtgg ccaccagcgt
     1261 ggcacctgag acaaaggagt gtccacctcc accacctgag acccctgccc tgggatgcct
     1321 ggcagcatgt ggaaggacaa tccagcggca cctggccttt gacatcctgc ggcacaacac
     1381 cggctactgc gtgtatatcc tgggcgtgat gtggtctgtg ctgggcttcc ctctgccaca
     1441 ggtgtttctg gtgccatacg ccatgtggca cagcgtggat gagcagcagg ccgccctgct
     1501 gatcagcatc atcggcttct ccaatatctt tctgagacca ctggcaggac tgatggcagg
     1561 cagacccgcc ttcgcctctc acaggaagta tctgtttagc ctggccctgc tgctgaacgg
     1621 cctgacaaat ctggtgtgcg cagcatccgg cgacttctgg gtgctggtgg gctactgtct
     1681 ggcctatagc gtgtccatgt ctggcatcgg cgccctgatc tttcaggtgc tgatggacat
     1741 cgtgccaatg gatcagttcc ccagggccct gggcctgttt accgtgctgg acggcctggc
     1801 cttcctgatc agcccaccac tggcaggact gctgctggat gccacaaaca atttctccta
     1861 cgtgttttat atgtcctctt tctttctgat ctccgccgcc ctgttcatgg gcggctcttt
     1921 ttatgccctg cagaagaagg agcagggcaa gcaggcagtg gcagcagacg ccctggagag
     1981 agatctgttc ctggaggcca aggatggccc tggcaagcag aggagcccag agatcatgtg
     2041 ccagagctcc aggcagccaa ggcctgcagg cgtgaacaag cacctgtggg gatgtccagc
     2101 atctagccgg accagccacg agtggctgct gtggcctaag gccgtgctgc aggcaaagca
     2161 gaccgccctg ggatggaatt cccccacaga cccagctttc ttgtacaaag ttggcattat
     2221 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2281 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2341 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2401 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2461 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2521 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2581 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2641 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2701 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2761 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2821 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2881 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2941 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3001 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3061 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3121 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3181 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3241 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3301 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3361 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3421 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3481 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3541 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3601 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3661 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3721 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3781 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3841 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3901 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3961 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4021 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
