LOCUS       sequence-260980-        4527 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 4527)
  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 4527)
  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_SLC5A4
FEATURES             Location/Qualifiers
     source          1..4527
                     /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(2652..2751)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2768..2787)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2769..2787)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2792..2808)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2792..2808)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2910..2929)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2921..3730
                     /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(2988..3007)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3877..4465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4366..4385
                     /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 accatggcct ccaccgtgag cccatccaca atcgcagaga ccccagagcc
      721 acctccactg tccgaccaca tcagaaacgc cgccgatatc tctgtgatcg tgatctactt
      781 cctggtggtc atggcagtgg gactgtgggc catgctgaag acaaatagag gcaccatcgg
      841 cggcttcttt ctggcaggaa gggatatggc atggtggcca atgggcgcct ctctgtttgc
      901 cagcaacatc ggctccaatc actatgtggg actggcagga acaggagcag caagcggcgt
      961 ggccacagtg accttcgagt ggaccagctc cgtgatgctg ctgatcctgg gctggatctt
     1021 tgtgccaatc tacatcaagt ccggcgtgat gaccatgccc gagtacctta aaaagcgctt
     1081 cggaggagag cgcctgcagg tgtacctgag catcctgtcc ctgtttatct gcgtggtgct
     1141 gctgatcagc gccgacatct tcgccggcgc catctttatc aagctggccc tgggcctgga
     1201 tctgtatctg gccatcttca tcctgctggc catgacagcc gtgtacacca caaccggagg
     1261 actggcatct gtgatctata cagacaccct gcagaccatc atcatgctga tcggcagctt
     1321 catcctgatg ggcttcgcct ttaacgaagt gggcggctac gagtctttta cagagaagta
     1381 tgtgaacgcc acccctagcg tggtggaggg cgacaatctg acaatctctg ccagctgcta
     1441 caccccacgc gccgattcct tccacatctt tcgggacgca gtgaccggcg atatcccatg
     1501 gccaggcatc atcttcggca tgcccatcac agccctgtgg tattggtgta ccaatcaggt
     1561 catcgtgcag cggtgcctgt gcggcaagga tatgagccac gtgaaggccg cctgcatcat
     1621 gtgcgcctac ctgaagctgc tgcctatgtt tctgatggtc atgccaggca tgatcagccg
     1681 gatcctgtat accgacatgg tggcctgcgt ggtgccttcc gagtgcgtga agcactgtgg
     1741 cgtggatgtg ggctgtacaa actacgccta tcccaccatg gtgctggagc tgatgcctca
     1801 gggcctgaga ggcctgatgc tgtccgtgat gctggcctct ctgatgtcta gcctgacatc
     1861 catcttcaat tccgcctcta cactgtttac catcgacctg tacaccaaga tgcgcaagca
     1921 ggcctctgag aaggagctgc tgatcgccgg ccggatcttc gtgctgctgc tgacagtggt
     1981 gagcatcgtg tgggtgcccc tggtgcaggt gtcccagaac ggccagctga tccactacac
     2041 cgagtccatc tcctcttatc tgggaccacc tatcgcagcc gtgttcgtgc tggccatctt
     2101 ttgcaagaga gtgaatgagc agggagcatt ctggggactg atggtgggac tggcaatggg
     2161 actgatcagg atgatcacag agtttgcata cggaaccgga agctgtctgg caccatccaa
     2221 ctgccctaag atcatctgtg gcgtgcacta cctgtatttt tctatcgtgc tgttctttgg
     2281 cagcatgctg gtgacactgg gcatcagcct gctgaccaag cctatcccag acgtgcacct
     2341 gtaccggctg tgctgggtgc tgagaaattc cacagaggag aggatcgaca tcgatgccga
     2401 ggagaagtct caggaggaga ccgacgatgg cgtggaggag gactaccccg agaagtctag
     2461 gggctgcctg aagaaggcct atgatctgtt ctgtggcctg cagaagggcc ctaagctgac
     2521 aaaggaggag gaggaggccc tgagcaagaa gctgacagat acctctgagc ggccaagctg
     2581 gagaaccatc gtgaacatca atgccatcct gctgctggcc gtggtggtgt ttatccacgg
     2641 ctactatgcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2701 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2761 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2821 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2881 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2941 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3001 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3061 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3121 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3181 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3241 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3301 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3361 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3421 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3481 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3541 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3601 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3661 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3721 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3781 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3841 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3901 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3961 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4021 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4081 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4141 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4201 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4261 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4321 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4381 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4441 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4501 cttttgctgg ccttttgctc acatgtt
//
