LOCUS       sequence-261099-        4479 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 4479)
  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 4479)
  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_SLC5A5
FEATURES             Location/Qualifiers
     source          1..4479
                     /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(2604..2703)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2720..2739)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2721..2739)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2744..2760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2862..2881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2873..3682
                     /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(2940..2959)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3829..4417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4318..4337
                     /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 accatggagg ccgtggagac aggagagagg cccaccttcg gcgcatggga
      721 ctacggcgtg tttgccctga tgctgctggt gtccacaggc atcggactgt gggtgggact
      781 ggcaagagga ggccagaggt ctgccgagga tttctttacc ggaggccgga gactggccgc
      841 cctgcctgtg ggcctgtccc tgtctgccag cttcatgagc gccgtgcagg tgctgggagt
      901 gccatccgag gcataccgct atggcctgaa gtttctgtgg atgtgcctgg gccagctgct
      961 gaattccgtg ctgacagccc tgctgttcat gccagtgttt taccggctgg gcctgacctc
     1021 cacatacgag tatctggaga tgaggttctc tagggccgtg cggctgtgcg gcaccctgca
     1081 gtatatcgtg gccacaatgc tgtacaccgg catcgtgatc tatgcccccg ccctgatcct
     1141 gaaccaggtg acaggcctgg acatctgggc cagcctgctg tccaccggca tcatctgcac
     1201 cttttacaca gcagtgggag gaatgaaggc agtggtgtgg acagacgtgt tccaggtggt
     1261 ggtcatgctg tccggatttt gggtggtgct ggcaaggggc gtgatgctgg tgggaggacc
     1321 ccggcaggtg ctgaccctgg cccagaacca cagcagaatc aatctgatgg acttcaaccc
     1381 tgatccaagg tcccgctata ccttctggac atttgtggtg ggcggcacac tggtgtggct
     1441 gagcatgtac ggcgtgaatc aggcacaggt gcagcggtac gtggcatgtc ggaccgagaa
     1501 gcaggccaag ctggccctgc tgatcaacca agtgggcctg tttctgatcg tgagctccgc
     1561 cgcctgctgt ggcatcgtga tgttcgtgtt ttacaccgac tgcgatcctc tgctgctggg
     1621 cagaatctcc gcccccgacc agtatatgcc tctgctggtg ctggacatct tcgaggatct
     1681 gccaggagtg cctggactgt ttctggcatg tgcctactct ggcaccctga gcacagcctc
     1741 cacctctatc aatgccatgg ccgccgtgac cgtggaggat ctgatcaagc ctcggctgag
     1801 aagcctggcc ccaaggaagc tggtcatcat ctccaagggc ctgtctctga tctatggatc
     1861 cgcctgcctg acagtggccg ccctgtctag cctgctggga ggaggcgtgc tgcagggctc
     1921 cttcaccgtg atgggcgtga tctctggacc tctgctggga gccttcatcc tgggaatgtt
     1981 tctgccagcc tgtaacaccc caggcgtgct ggcaggactg ggagcaggcc tggccctgtc
     2041 tctgtgggtg gccctgggag ccacactgta cccacctagc gagcagacca tgagagtgct
     2101 gccatcctct gccgcaagat gcgtggccct gagcgtgaat gcatccggac tgctggaccc
     2161 agccctgctg ccagccaacg atagctccag agccccatct agcggaatgg acgcaagcag
     2221 gcccgccctg gccgattctt tctacgccat cagctatctg tactatggcg ccctgggcac
     2281 cctgaccaca gtgctgtgcg gcgccctgat ctcttgtctg acaggaccaa ccaagcgcag
     2341 cacactggca cctggactgc tgtggtggga cctggcaagg cagaccgcat ctgtggcacc
     2401 aaaggaggag gtggccatcc tggacgataa tctggtgaag ggccctgagg agctgccaac
     2461 aggcaataag aagccacccg gctttctgcc caccaacgag gatagactgt tctttctggg
     2521 acagaaggag ctggagggag caggaagctg gacaccttgc gtgggacacg acggaggaag
     2581 ggatcagcag gagaccaacc tggacccagc tttcttgtac aaagttggca ttataagaaa
     2641 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2701 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2761 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2821 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2881 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2941 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3001 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3061 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3121 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3181 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3241 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3301 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3361 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3421 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3481 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3541 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3601 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3661 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3721 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3781 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4441 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
