LOCUS       sequence-261924-        4380 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 4380)
  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 4380)
  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_SLC5A8
FEATURES             Location/Qualifiers
     source          1..4380
                     /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(2505..2604)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2621..2640)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2622..2640)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2645..2661)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2645..2661)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2763..2782)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2774..3583
                     /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(2841..2860)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3730..4318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4219..4238
                     /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 accatggaca cacctagggg catcggcacc ttcgtggtgt gggattacgt
      721 ggtgtttgcc ggcatgctgg tcatctctgc cgccatcggg atctactacg cattcgctgg
      781 tggaggacag cagaccagca aggactttct gatgggcggc cggagaatga cagcagtgcc
      841 tgtggccctg agcctgaccg catctttcat gagcgccgtg acagtgctgg gaaccccatc
      901 cgaggtgtac cgcttcggcg ccatcttttc tatcttcgcc tttacctatt tctttgtggt
      961 ggtcatcagc gccgaggtgt tcctgcccgt gttttacaag ctgggcatca catccaccta
     1021 cgagtatctg gagctgcggt tcaataagtg cgtgagactg tgcggcaccg tgctgtttat
     1081 cgtgcagaca atcctgtaca ccggcatcgt gatctatgcc cctgccctgg ccctgaacca
     1141 ggtgacaggc ttcgacctgt ggggagcagt ggtggcaacc ggcgtggtgt gcaccttcta
     1201 ctgtaccctg ggcggcctga aggccgtgat ctggacagac gtgttccaag tgggcatcat
     1261 ggtggccggc tttgcctctg tgatcatcca ggccgtggtc atgcagggcg gcatcagcac
     1321 catcctgaat gacgcctatg atggcggcag gctgaacttc tggaacttta atccaaaccc
     1381 cctgcagcgc cacaccttct ggacaatcat catcggcggc accttcacct ggacaagcat
     1441 ctacggcgtg aatcagtccc aggtgcagag gtatatctcc tgcaagtctc gcttccaggc
     1501 caagctgagc ctgtacatca acctggtggg cctgtgggcc atcctgacat gcagcgtgtt
     1561 ctgcggcctg gccctgtaca gccggtatca cgactgcgat ccatggaccg ccaagaaggt
     1621 gtccgcccca gaccagctga tgccttacct ggtgctggac atcctgcagg attatcctgg
     1681 actgccagga ctgttcgtgg catgtgcata ttctggcacc ctgagcacag tgagctcctc
     1741 tatcaatgca ctggcagcag tgaccgtgga ggatctgatc aagccctact ttaggagcct
     1801 gtccgagcgg agcctgagct ggatctccca gggcatgtct gtggtgtatg gcgccctgtg
     1861 catcggaatg gccgccctgg caagcctgat gggcgccctg ctgcaggccg ccctgtccgt
     1921 gttcggaatg gtgggaggac cactgatggg actgttcgca ctgggaatcc tggtgccttt
     1981 tgccaattcc atcggcgccc tggtgggact gatggcaggc tttgccatct ctctgtgggt
     2041 gggaatcgga gcacagatct acccacctct gccagagcgg acactgccac tgcacctgga
     2101 catccagggc tgtaactcca catataatga gaccaacctg atgaccacaa ccgagatgcc
     2161 cttcaccacc agcgtgttcc agatctacaa tgtgcagaga acccccctga tggacaactg
     2221 gtattccctg tcttacctgt atttcagcac cgtgggcaca ctggtgaccc tgctggtggg
     2281 catcctggtg agcctgtcca caggcggccg gaagcagaat ctggatccca gatacatcct
     2341 gaccaaggag gacttcctgt ccaacttcga tatcttcaag aagaagaagc acgtgctgag
     2401 ctataagtcc caccctgtgg aggacggcgg cacagataat ccagccttta accacatcga
     2461 gctgaattct gatcagagcg gcaagtccaa cggcaccagg ctggacccag ctttcttgta
     2521 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2581 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2641 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2701 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2761 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2821 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2881 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2941 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3001 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3061 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3121 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3181 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3241 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3301 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3361 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3421 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3481 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3541 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3601 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3661 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3721 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3781 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4201 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4261 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4321 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
