LOCUS       sequence-263874-        4566 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 4566)
  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 4566)
  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_SLC5A2
FEATURES             Location/Qualifiers
     source          1..4566
                     /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(2691..2790)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2807..2826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2808..2826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2831..2847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2831..2847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2949..2968)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2960..3769
                     /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(3027..3046)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3916..4504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4405..4424
                     /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 agcacaccga ggcaggaagc gcccctgaga tgggcgccca
      721 gaaggccctg atcgataacc cagccgacat cctggtcatc gccgcctact tcctgcttgt
      781 tataggcgtg gggctctggt ctatgtgcag aaccaatagg ggcacagtgg gcggctattt
      841 tctggccggc agatccatgg tgtggtggcc agtgggagca agcctgttcg cctccaacat
      901 cggatctgga cactttgtgg gactggcagg aaccggagca gcatccggcc tggccgtggc
      961 cggcttcgag tggaatgccc tgtttgtggt gctgctgctg ggatggctgt tcgcaccagt
     1021 gtacctgacc gccggcgtga tcacaatgcc ccagtatctg aggaagcggt tcggcggccg
     1081 gagaatcagg ctgtacctgt ccgtgctgtc tctgttcctg tatatcttta caaagatcag
     1141 cgtggatatg ttctccggcg ccgtgtttat ccagcaggcc ctgggctgga acatctacgc
     1201 cagcgtgatc gccctgctgg gcatcaccat gatctacacc gtgacaggag gactggccgc
     1261 cctgatgtat accgacacag tgcagacatt cgtgatcctg ggcggcgcct gcatcctgat
     1321 gggctatgca ttccacgaag tgggaggata cagcggactg tttgataagt atctgggagc
     1381 agccaccagc ctgacagtgt ccgaggaccc agcagtgggc aatatcagct ccttttgtta
     1441 ccggccaaga cccgattcct atcacctgct gaggcaccct gtgaccggcg acctgccttg
     1501 gccagccctg ctgctgggac tgacaatcgt gtctggctgg tactggtgca gcgatcaggt
     1561 catcgtgcag cggtgtctgg ccggcaagag cctgacccac atcaaggccg gctgcatcct
     1621 gtgcggctac ctgaagctga caccaatgtt cctgatggtc atgcccggca tgatctctcg
     1681 catcctgtat cctgacgagg tggcatgcgt ggtgccagag gtgtgcaggc gggtgtgcgg
     1741 aaccgaagtg ggctgtagca acatcgccta ccccagactg gtggtgaagc tgatgcctaa
     1801 tggcctgagg ggactgatgc tggccgtgat gctggccgcc ctgatgtcta gcctggcctc
     1861 catcttcaac tcctctagca ccctgtttac aatggatatc tatacaaggc tgaggcctag
     1921 ggcaggcgac agggagctgc tgctggtggg ccggctgtgg gtggtgttca tcgtggtggt
     1981 gtccgtggcc tggctgccag tggtgcaggc agcacaggga ggccagctgt ttgattacat
     2041 ccaggccgtg tcctcttatc tggcaccacc cgtgagcgcc gtgttcgtgc tggccctgtt
     2101 tgtgccaaga gtgaatgagc agggagcatt ctggggactg atcggaggac tgctgatggg
     2161 actggcaagg ctgatccccg agttctcttt tggctctgga agctgcgtgc agcctagcgc
     2221 ctgcccagcc ttcctgtgcg gagtgcacta cctgtatttt gccatcgtgc tgttcttttg
     2281 cagcggcctg ctgaccctga cagtgtccct gtgcaccgca ccaatcccta ggaagcacct
     2341 gcaccggctg gtgttctctc tgagacacag caaggaggag agggaggacc tggatgcaga
     2401 cgagcagcag ggaagctccc tgccagtgca gaacggatgt cctgagtccg ccatggagat
     2461 gaatgagcca caggcaccag caccatctct gttccgccag tgcctgctgt ggttttgtgg
     2521 catgtccagg ggaggagtgg gatctccacc acctctgacc caggaggagg cagcagcagc
     2581 agcacggaga ctggaggata tctccgagga cccttcttgg gccagggtgg tgaacctgaa
     2641 tgccctgctg atgatggccg tggccgtgtt cctgtggggc ttttacgccg acccagcttt
     2701 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2761 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2821 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2881 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2941 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     3001 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     3061 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     3121 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     3181 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     3241 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     3301 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3361 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3421 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3481 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3541 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3601 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3661 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3721 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3781 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3841 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3901 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3961 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4021 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4081 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4141 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4201 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4261 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4321 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4381 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4441 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4501 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4561 catgtt
//
