LOCUS       sequence-263085-        3438 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 3438)
  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 3438)
  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-TMEM241
FEATURES             Location/Qualifiers
     source          1..3438
                     /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(1563..1662)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1679..1698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1680..1698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1703..1719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1703..1719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1821..1840)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1832..2641
                     /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(1899..1918)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2788..3376
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3277..3296
                     /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 accatgtgcg tgcggagaag cctggtgggc ctgaccttct gcacatgtta
      721 cctggcctcc tatctgacca acaagtacgt gctgtctgtg ctgaagttca cctatccaac
      781 actgtttcag ggctggcaga cactgatcgg cggcctgctg ctgcacgtgt cctggaagct
      841 gggctgggtg gagatcaata gctcctctcg gagccacgtg ctggtgtggc tgcctgcatc
      901 cgtgctgttc gtgggcatca tctacgcagg atccagggcc ctgtctaggc tggcaatccc
      961 agtgttcctg accctgcaca acgtggccga agtgatcatc tgcggctatc agaagtgttt
     1021 tcagaaggag aagacatctc ccgccaagat ctgcagcgcc ctgctgctgc tggcagcagc
     1081 aggctgtctg cccttcaacg actcccagtt taatcctgat ggctacttct gggccatcat
     1141 ccacctgctg tgcgtgggcg cctataagat cctgcagaag tctcagaagc cttctgccct
     1201 gagcgacatc gatcagcagt acctgaacta tatcttctcc gtggtgctgc tggcctttgc
     1261 ctctcaccca accggcgacc tgtttagcgt gctggatttc ccctttctgt acttctatcg
     1321 gtttcacgga agctgctgtg catccggctt cctgggcttc tttctgatgt ttagcacagt
     1381 gaagctgaag aatctgctgg caccaggaca gtgcgcagca tggatcttct ttgccaagat
     1441 catcaccgcc ggcctgtcta tcctgctgtt cgacgccatc ctgacaagcg ccaccacagg
     1501 atgtctgctg ctgggcgccc tgggagaggc cctgctggtg ttttccgaga ggaagagctc
     1561 cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1621 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1681 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1741 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1801 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1861 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     1921 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     1981 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2041 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2101 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2161 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2221 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2281 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2341 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2401 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2461 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2521 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2581 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2641 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2701 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3421 gccttttgct cacatgtt
//
