LOCUS       sequence-261843-        4221 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 4221)
  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 4221)
  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_SLC22A5
FEATURES             Location/Qualifiers
     source          1..4221
                     /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(2346..2445)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2462..2481)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2463..2481)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2486..2502)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2486..2502)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2604..2623)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2615..3424
                     /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(2682..2701)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3571..4159
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4060..4079
                     /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 accatgcgcg actacgatga ggtgaccgcc ttcctgggag agtggggacc
      721 atttcagcgg ctgatcttct ttctgctgtc tgccagcatc atccctaacg gcttcaccgg
      781 cctgagctcc gtgtttctga tcgccacacc agagcacagg tgcagagtgc cagacgcagc
      841 aaacctgtct agcgcctgga gaaatcacac cgtgcctctg aggctgaggg atggaaggga
      901 ggtgccacac tcctgtcgga gataccggct ggccacaatc gccaatttct ctgccctggg
      961 actggagcct ggaagggacg tggatctggg ccagctggag caggagagct gcctggacgg
     1021 ctgggagttt tctcaggacg tgtacctgag caccatcgtg acagagtgga atctggtgtg
     1081 cgaagacgac tggaaagccc cactgaccat ctctctgttc tttgtgggcg tgctgctggg
     1141 ctccttcatc tctggccagc tgagcgaccg gttcggcaga aagaatgtgc tgtttgtgac
     1201 catgggcatg cagacaggct tctcctttct gcagatcttc tctaagaact tcgagatgtt
     1261 tgtggtgctg tttgtgctgg tgggcatggg ccagatctcc aattacgtgg ccgccttcgt
     1321 gctgggcacc gagatcctgg gcaagagcgt gagaatcatc ttttccacac tgggcgtgtg
     1381 catcttctac gcctttggct atatggtgct gccactgttc gcctacttca tcagggactg
     1441 gcgcatgctg ctggtggccc tgaccatgcc aggcgtgctg tgcgtggccc tgtggtggtt
     1501 catccctgag tctccaaggt ggctgatcag ccagggccgc tttgaggagg ccgaagtgat
     1561 catccggaag gcagcaaagg caaacggaat cgtggtgccc tccacaatct tcgacccttc
     1621 tgagctgcag gatctgtcct ctaagaagca gcagagccac aacatcctgg atctgctgcg
     1681 gacctggaat atcagaatgg tgacaatcat gtccatcatg ctgtggatga ccatcagcgt
     1741 gggctacttt ggcctgtccc tggacacacc caacctgcac ggcgatatct tcgtgaattg
     1801 ctttctgagc gccatggtgg aggtgcccgc ctatgtgctg gcctggctgc tgctgcagta
     1861 cctgcctagg cgctattcta tggcaaccgc cctgttcctg ggaggaagcg tgctgctgtt
     1921 tatgcagctg gtgccccctg atctgtacta tctggccacc gtgctggtca tggtgggcaa
     1981 gttcggcgtg acagccgcct tttctatggt gtacgtgtat accgccgagc tgtaccccac
     2041 agtggtgaga aatatgggag tgggcgtgag ctccaccgca tccaggctgg gaagcatcct
     2101 gtccccatac ttcgtgtatc tgggcgccta cgaccgcttt ctgccctata tcctgatggg
     2161 cagcctgacc atcctgacag ccatcctgac actgttcctg cctgagtcct ttggcacccc
     2221 actgcccgac acaatcgatc agatgctgcg ggtgaagggc atgaagcaca gaaagacccc
     2281 ttcccacaca aggatgctga aggatggcca ggagcggcca accatcctga agtctacagc
     2341 cttcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2401 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2461 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2521 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2581 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2641 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2701 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2761 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2821 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2881 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2941 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3001 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3061 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3121 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3181 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3241 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3301 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3361 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3421 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3481 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3541 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3601 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3661 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3721 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3781 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3841 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3901 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3961 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4021 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4081 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4141 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4201 ctggcctttt gctcacatgt t
//
