LOCUS       sequence-352162-        4569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4569)
  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 4569)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2126..2225)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2242..2261)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2243..2261)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2266..2282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2266..2282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2384..2403)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2395..3204
                     /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(2462..2481)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3351..3939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3840..3859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4093..4110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4269..4296)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4388..4474)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4523..4545
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4537..4554
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4538..4554
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga ggagcacacc
      121 gaggcaggaa gcgcccctga gatgggcgcc cagaaggccc tgatcgataa cccagccgac
      181 atcctggtca tcgccgccta cttcctgctt gttataggcg tggggctctg gtctatgtgc
      241 agaaccaata ggggcacagt gggcggctat tttctggccg gcagatccat ggtgtggtgg
      301 ccagtgggag caagcctgtt cgcctccaac atcggatctg gacactttgt gggactggca
      361 ggaaccggag cagcatccgg cctggccgtg gccggcttcg agtggaatgc cctgtttgtg
      421 gtgctgctgc tgggatggct gttcgcacca gtgtacctga ccgccggcgt gatcacaatg
      481 ccccagtatc tgaggaagcg gttcggcggc cggagaatca ggctgtacct gtccgtgctg
      541 tctctgttcc tgtatatctt tacaaagatc agcgtggata tgttctccgg cgccgtgttt
      601 atccagcagg ccctgggctg gaacatctac gccagcgtga tcgccctgct gggcatcacc
      661 atgatctaca ccgtgacagg aggactggcc gccctgatgt ataccgacac agtgcagaca
      721 ttcgtgatcc tgggcggcgc ctgcatcctg atgggctatg cattccacga agtgggagga
      781 tacagcggac tgtttgataa gtatctggga gcagccacca gcctgacagt gtccgaggac
      841 ccagcagtgg gcaatatcag ctccttttgt taccggccaa gacccgattc ctatcacctg
      901 ctgaggcacc ctgtgaccgg cgacctgcct tggccagccc tgctgctggg actgacaatc
      961 gtgtctggct ggtactggtg cagcgatcag gtcatcgtgc agcggtgtct ggccggcaag
     1021 agcctgaccc acatcaaggc cggctgcatc ctgtgcggct acctgaagct gacaccaatg
     1081 ttcctgatgg tcatgcccgg catgatctct cgcatcctgt atcctgacga ggtggcatgc
     1141 gtggtgccag aggtgtgcag gcgggtgtgc ggaaccgaag tgggctgtag caacatcgcc
     1201 taccccagac tggtggtgaa gctgatgcct aatggcctga ggggactgat gctggccgtg
     1261 atgctggccg ccctgatgtc tagcctggcc tccatcttca actcctctag caccctgttt
     1321 acaatggata tctatacaag gctgaggcct agggcaggcg acagggagct gctgctggtg
     1381 ggccggctgt gggtggtgtt catcgtggtg gtgtccgtgg cctggctgcc agtggtgcag
     1441 gcagcacagg gaggccagct gtttgattac atccaggccg tgtcctctta tctggcacca
     1501 cccgtgagcg ccgtgttcgt gctggccctg tttgtgccaa gagtgaatga gcagggagca
     1561 ttctggggac tgatcggagg actgctgatg ggactggcaa ggctgatccc cgagttctct
     1621 tttggctctg gaagctgcgt gcagcctagc gcctgcccag ccttcctgtg cggagtgcac
     1681 tacctgtatt ttgccatcgt gctgttcttt tgcagcggcc tgctgaccct gacagtgtcc
     1741 ctgtgcaccg caccaatccc taggaagcac ctgcaccggc tggtgttctc tctgagacac
     1801 agcaaggagg agagggagga cctggatgca gacgagcagc agggaagctc cctgccagtg
     1861 cagaacggat gtcctgagtc cgccatggag atgaatgagc cacaggcacc agcaccatct
     1921 ctgttccgcc agtgcctgct gtggttttgt ggcatgtcca ggggaggagt gggatctcca
     1981 ccacctctga cccaggagga ggcagcagca gcagcacgga gactggagga tatctccgag
     2041 gacccttctt gggccagggt ggtgaacctg aatgccctgc tgatgatggc cgtggccgtg
     2101 ttcctgtggg gcttttacgc ctaagaccca gctttcttgt acaaagttgg cattataaga
     2161 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     2221 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     2281 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     2341 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     2401 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2461 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2521 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2581 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2641 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2701 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2761 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2821 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2881 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2941 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     3001 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     3061 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3121 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3181 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3241 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3301 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4141 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4201 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     4261 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     4321 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     4381 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     4441 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4501 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     4561 ctcgggccc
//
