LOCUS       sequence-479325-        4578 bp    DNA     circular SYN 07-AUG-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 4578)
  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 4578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 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-SLC5A11_STOP
FEATURES             Location/Qualifiers
     source          1..4578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     455..472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(631..658)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(750..836)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     885..907
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     899..916
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     900..916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    932..1031
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3066..3165)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3182..3201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3183..3201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3206..3222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3206..3222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3324..3343)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3335..4144
                     /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(3402..3421)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(4291..4578,1..301)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
       61 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      481 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      541 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
      601 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
      661 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      721 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      781 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
      841 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
      901 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
      961 gacctgttcg ttgcaacaca ttgatgagca atgctttttt ataatgccaa ctttgtacaa
     1021 aaaagcaggc tccaccatgg agtccggcac cagctcccct cagccacctc agctggaccc
     1081 actggatgca ttccctcaga agggcctgga gccaggcgat atcgccgtgc tggtgctgta
     1141 cttcctgttt gtgctggcag tgggactgtg gagcaccgtg aagacaaagc gggacaccgt
     1201 gaagggctat tttctggcag gaggcgatat ggtgtggtgg ccagtgggag catctctgtt
     1261 cgccagcaac gtgggatccg gacactttat cggactggca ggatctggag cagcaacagg
     1321 aatctccgtg tctgcctacg agctgaatgg cctgttcagc gtgctgatgc tggcctggat
     1381 ctttctgcca atttatatcg ccggccaggt gaccacaatg cccgagtatc tgcggaagag
     1441 attcggcggc atcaggatcc ctatcatcct ggccgtgctg tacctgttca tctatatctt
     1501 taccaagatc agcgtggaca tgtacgccgg cgccatcttt atccagcagt ccctgcacct
     1561 ggatctgtat ctggcaatcg tgggactgct ggcaatcacc gccgtgtaca cagtggcagg
     1621 aggactggca gccgtgatct ataccgacgc cctgcagaca ctgatcatgc tgatcggcgc
     1681 cctgaccctg atgggatact ccttcgccgc agtgggagga atggagggcc tgaaggagaa
     1741 gtattttctg gccctggcct ctaaccggag cgagaattct agctgcggac tgcctaggga
     1801 ggacgcattc cacatctttc gcgacccact gacaagcgat ctgccatggc ccggcgtgct
     1861 gttcggcatg tccatccctt ctctgtggta ctggtgtacc gatcaggtca tcgtgcagag
     1921 gacactggcc gccaagaacc tgtctcacgc aaagggaggc gccctgatgg cagcatatct
     1981 gaaggtgctg ccactgttca tcatggtgtt tcccggcatg gtgagccgca tcctgtttcc
     2041 tgaccaggtg gcctgcgccg atccagagat ctgccagaag atctgtagca atccatccgg
     2101 ctgttctgac atcgcctacc ccaagctggt gctggagctg ctgcctaccg gactgagggg
     2161 actgatgatg gccgtgatgg tggccgccct gatgtcctct ctgacaagca tcttcaacag
     2221 cgcctccacc atctttacaa tggatctgtg gaatcacctg aggccaaggg catccgagaa
     2281 ggagctgatg atcgtgggca gagtgttcgt gctgctgctg gtgctggtgt ccatcctgtg
     2341 gatccctgtg gtgcaggcat ctcagggagg acagctgttt atctacatcc agtccatcag
     2401 ctcctatctg cagccacccg tggccgtggt gttcatcatg ggctgctttt ggaagcggac
     2461 caacgagaag ggagcattct ggggactgat ctccggactg ctgctgggac tggtgagact
     2521 ggtgctggac ttcatctacg tgcagccccg gtgtgaccag cctgatgaga gaccagtgct
     2581 ggtgaagtct atccactacc tgtatttctc tatgatcctg agcaccgtga cactgatcac
     2641 cgtgagcaca gtgtcctggt tcacagagcc tccatctaag gagatggtga gccacctgac
     2701 ctggtttaca aggcacgacc cagtggtgca gaaggagcag gcaccacctg cagcaccact
     2761 gtccctgacc ctgtctcaga acggcatgcc tgaggcctct agctcctcta gcgtgcagtt
     2821 cgagatggtg caggagaata ccagcaagac acactcctgc gatatgacac ccaagcagag
     2881 caaggtggtg aaggccatcc tgtggctgtg cggcatccag gagaagggca aggaggagct
     2941 gccagcaagg gcagaggcaa tcatcgtgtc cctggaggag aaccccctgg tgaagaccct
     3001 gctggacgtg aatctgatct tctgcgtgtc ttgtgccatc ttcatctggg gctactttgc
     3061 ctaagaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     3121 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3181 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3241 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     3301 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     3361 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     3421 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     3481 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3541 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3601 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3661 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3721 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3781 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3841 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3901 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3961 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4021 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4081 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4141 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4201 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4561 gttggactca agacgata
//
