LOCUS       sequence-263064-        4767 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 4767)
  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 4767)
  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-LETM1
FEATURES             Location/Qualifiers
     source          1..4767
                     /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(2892..2991)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3008..3027)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3009..3027)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3032..3048)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3032..3048)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3150..3169)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3161..3970
                     /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(3228..3247)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4117..4705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4606..4625
                     /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 accatggcaa gcatcctgct gcggtcctgc aggggaagag caccagcaag
      721 actgccacct ccaccaaggt acaccgtgcc aagaggaagc ccaggcgacc cagcccacct
      781 gtcttgcgcc agcacactgg gcctgaggaa ctgtctgaat gtgcccttcg gctgctgtac
      841 ccctatccac ccagtgtata caagctccag gggcgatcac ctgggatgct gggcactgag
      901 gccagagtgt ctgaggatcg tgtcccgcgc cccttggacc tccacatctg tgggctttgt
      961 ggcagtggga ccacagtgtc tgcccgtgcg cggatggcac tctagcaggc cagtgagaga
     1021 cgattctgtg gtggagaagt ccctgaagtc tctgaaggac aagaacaaga agctggagga
     1081 gggaggaccc gtgtacagcc ctccagccga ggtggtggtg aagaagtccc tgggacagcg
     1141 ggtgctggac gagctgaagc actactatca cggcttcaga ctgctgtgga tcgataccaa
     1201 gatcgccgcc cggatgctgt ggagaatcct gaatggccac agcctgacac ggagagagag
     1261 gcgccagttc ctgaggatct gcgccgatct gtttcgcctg gtgcctttcc tggtgtttgt
     1321 ggtggtgcct ttcatggagt ttctgctgcc agtggccgtg aagctgttcc ctaacatgct
     1381 gccatccacc tttgagacac agtctctgaa ggaggagcgg ctgaagaagg agctgagagt
     1441 gaagctggag ctggccaagt tcctgcagga caccatcgag gagatggccc tgaagaataa
     1501 ggccgccaag ggcagcgcca ccaaggattt ttccgtgttc tttcagaaga tcagggagac
     1561 aggagagagg ccatccaacg aggagatcat gaggttctct aagctgtttg aggacgagct
     1621 gaccctggat aatctgacac gccctcagct ggtggccctg tgcaagctgc tggagctgca
     1681 gtccatcggc accaacaatt tcctgaggtt tcagctgaca atgaggctgc gctccatcaa
     1741 ggccgacgat aagctgatcg ccgaggaggg cgtggactct ctgaacgtga aggagctgca
     1801 ggcagcatgt agggcaagag gaatgcgggc cctgggcgtg accgaggaca ggctgagggg
     1861 acagctgaag cagtggctgg atctgcacct gcaccaggag atccctacct ctctgctgat
     1921 cctgagcaga gccatgtatc tgcccgacac actgtctcct gccgatcagc tgaagagcac
     1981 cctgcagaca ctgccagaga tcgtggccaa ggaggcacag gtgaaggtgg cagaggtgga
     2041 gggagagcag gtggacaata aggccaagct ggaggccacc ctgcaggagg aggcagcaat
     2101 ccagcaggag caccgggaga aggagctgca gaagagatct gaggtggcca aggattttga
     2161 gcccgagagg gtggtggcag caccacagag gcctggaacc gagccccagc ctgagatgcc
     2221 tgacacagtg ctgcagagcg agaccctgaa ggatacagcc ccagtgctgg agggcctgaa
     2281 ggaggaggag atcaccaagg aggagatcga catcctgtct gatgcctgta gcaagctgca
     2341 ggagcagaag aagagcctga caaaggagaa ggaggagctg gagctgctga aggaggacgt
     2401 gcaggattac tctgaggacc tgcaggagat caagaaggag ctgagcaaga ccggcgagga
     2461 gaagtatgtg gaggagagca aggcctccaa gaggctgaca aagcgcgtgc agcagatgat
     2521 cggccagatc gacggcctga tcagccagct ggagatggat cagcaggcag gcaagctggc
     2581 accagcaaac ggaatgccaa ccggcgagaa tgtgatctcc gtggccgagc tgatcaacgc
     2641 catgaagcag gtgaagcaca tccccgagtc caagctgaca tctctggccg ccgccctgga
     2701 cgagaacaag gatggcaagg tcaatatcga cgatctggtg aaagtgatcg agctggtgga
     2761 caaggaggat gtgcacatca gcacctccca ggtggccgag atcgtggcca cactggagaa
     2821 ggaggagaag gtggaggaga aggagaaggc caaggagaag gccgagaagg aggtggccga
     2881 ggtgaagtcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2941 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     3001 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3061 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3121 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3181 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3241 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3301 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3361 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3421 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3481 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3541 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3601 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3661 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3721 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3781 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3841 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3901 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3961 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     4021 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4081 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4141 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4201 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4261 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4321 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4381 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4441 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4501 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4561 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4621 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4681 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4741 cttttgctgg ccttttgctc acatgtt
//
