LOCUS       sequence-261162-        4542 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 4542)
  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 4542)
  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_SLC5A1
FEATURES             Location/Qualifiers
     source          1..4542
                     /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(2667..2766)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2783..2802)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2784..2802)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2807..2823)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2807..2823)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2925..2944)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2936..3745
                     /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(3003..3022)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3892..4480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4381..4400
                     /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 gctccacctg gtctcctaag accacagccg tgacccgccc
      721 agtggagaca cacgagctga tccggaacgc cgccgatatc agcatcatcg tgatctactt
      781 cgtggtggtc atggcagtgg gactgtgggc catgttttct accaatagag gcacagtggg
      841 cggcttcttt ctggcaggca ggagcatggt gtggtggcca atcggagcaa gcctgttcgc
      901 ctccaacatc ggatctggac actttgtggg actggcagga accggagcag catccggcat
      961 cgccatcggc ggcttcgagt ggaatgcact ggtgctggtg gtggtgctgg gatggctgtt
     1021 tgtgccaatc tacatcaagg ccggcgtggt gaccatgccc gagtatctga ggaagcggtt
     1081 cggaggacag aggatccagg tgtacctgag cctgctgtcc ctgctgctgt atatctttac
     1141 aaagatctct gccgacatct tcagcggcgc catctttatc aacctggccc tgggcctgaa
     1201 tctgtacctg gccatcttcc tgctgctggc catcaccgcc ctgtacacca tcacaggcgg
     1261 cctggccgcc gtgatctata ccgacacact gcagaccgtg atcatgctgg tgggctccct
     1321 gatcctgaca ggcttcgcct ttcacgaagt gggcggctac gatgccttca tggagaagta
     1381 tatgaaggcc atccccacaa tcgtgtccga cggcaacacc acatttcagg agaagtgcta
     1441 cacccctaga gccgattctt tccacatctt tagggaccca ctgacaggcg atctgccatg
     1501 gcccggcttc atctttggca tgtctatcct gaccctgtgg tattggtgca cagatcaggt
     1561 catcgtgcag agatgtctgt ccgccaagaa tatgtctcac gtgaagggcg gctgcatcct
     1621 gtgcggctac ctgaagctga tgccaatgtt catcatggtc atgcccggca tgatcagcag
     1681 gatcctgtat accgagaaga tcgcctgcgt ggtgccttcc gagtgcgaga agtactgtgg
     1741 caccaaagtg ggctgtacaa acatcgccta tcccaccctg gtggtggagc tgatgcctaa
     1801 tggcctgcgc ggcctgatgc tgagcgtgat gctggcctcc ctgatgtcta gcctgacatc
     1861 catcttcaac tccgcctcta ccctgtttac aatggacatc tacgccaagg tgcggaagag
     1921 agccagcgag aaggagctga tgatcgccgg ccggctgttc atcctggtgc tgatcggcat
     1981 ctccatcgcc tgggtgccta tcgtgcagtc tgcccagagc ggccagctgt ttgattacat
     2041 ccagtccatc acctcttatc tgggaccacc tatcgcagcc gtgttcctgc tggccatctt
     2101 ttggaagagg gtgaacgagc caggagcatt ctggggactg atcctgggac tgctgatcgg
     2161 catcagccgg atgatcaccg agtttgccta tggcacaggc tcttgcatgg agcccagcaa
     2221 ttgccctaca atcatctgtg gcgtgcacta cctgtatttc gccatcatcc tgttcgccat
     2281 ctcctttatc accatcgtgg tcatctctct gctgacaaag cctatcccag acgtgcacct
     2341 gtacaggctg tgctggagcc tgcgcaactc caaggaggag cggatcgacc tggatgccga
     2401 ggaggagaat atccaggagg gccccaagga gaccatcgag atcgagacac aggtgcctga
     2461 gaagaagaag ggcatcttcc ggagagccta tgacctgttt tgcggactgg agcagcacgg
     2521 agcaccaaag atgaccgagg aggaggagaa ggccatgaag atgaagatga ccgatacaag
     2581 cgagaagccc ctgtggagaa ccgtgctgaa cgtgaatggc atcatcctgg tgacagtggc
     2641 cgtgttctgt cacgcctact ttgccgaccc agctttcttg tacaaagttg gcattataag
     2701 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2761 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2821 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2881 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2941 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     3001 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     3061 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3121 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3181 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3241 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3301 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3361 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3421 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3481 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3541 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3601 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3661 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3721 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3781 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3841 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3901 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3961 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4021 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4081 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4141 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4201 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4261 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4321 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4381 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4441 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4501 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
