LOCUS       sequence-259266-        2877 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 2877)
  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 2877)
  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_MPC1
FEATURES             Location/Qualifiers
     source          1..2877
                     /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(1002..1101)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1118..1137)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1119..1137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1142..1158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1142..1158)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1260..1279)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1271..2080
                     /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(1338..1357)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2227..2815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2716..2735
                     /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 accatggcag gcgccctggt gcggaaggca gcagactacg tgcggtctaa
      721 ggacttcaga gattatctga tgagcaccca cttttgggga ccagtggcaa actggggact
      781 gcctatcgcc gccatcaatg atatgaagaa gagcccagag atcatctccg gccggatgac
      841 cttcgccctg tgctgttact ccctgacatt catgaggttt gcctataagg tgcagccccg
      901 caactggctg ctgtttgcct gccacgccac aaatgaggtg gcacagctga tccagggagg
      961 caggctgatc aagcacgaga tgaccaagac agcctctgcc gacccagctt tcttgtacaa
     1021 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1081 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1141 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1201 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1261 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1321 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1381 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1441 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1501 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1561 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1621 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1681 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1741 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     1801 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     1861 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     1921 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     1981 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2041 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2101 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2161 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
