LOCUS       sequence-262982-        4038 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 4038)
  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 4038)
  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_TMEM104
FEATURES             Location/Qualifiers
     source          1..4038
                     /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(2163..2262)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2279..2298)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2280..2298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2303..2319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2303..2319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2421..2440)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2432..3241
                     /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(2499..2518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3388..3976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3877..3896
                     /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 accatggccg gcgagatcac agagaccggc gagctgtaca gctcctatgt
      721 gggcctggtg tacatgttta acctgatcgt gggaacaggc gccctgacca tgccaaaggc
      781 cttcgccacc gcaggatggc tggtgtccct ggtgctgctg gtgttcctgg gctttatgtc
      841 tttcgtgacc acaacctttg tgatcgaggc aatggcagca gcaaacgcgc agttgcattg
      901 gaaacgcatg gagaatctga aggaggagga ggacgatgac tctagcaccg ccagcgattc
      961 cgacgtgctg atcagagata actacgagag ggccgagaag cgccctatcc tgagcgtgca
     1021 gcggagaggc tcccctaatc catttgagat caccgaccgg gtggagatgg gccagatggc
     1081 cagcatgttc tttaacaagg tcggcgtgaa tctgttctac ttttgcatca tcgtgtacct
     1141 gtatggcgat ctggccatct atgccgccgc cgtgccattc tccctgatgc aggtgacatg
     1201 ctctgccacc ggcaacgata gctgtggcgt ggaggccgac acaaagtaca atgataccga
     1261 caggtgttgg ggaccactga ggagggtgga tgcatacaga atctatctgg ccatcttcac
     1321 actgctgctg ggccctttta ccttctttga cgtgcagaag acaaagtatc tgcagatcct
     1381 gaccagcctg atgaggtgga tcgccttcgc cgtgatgatc gtgctggccc tgatccgcat
     1441 cggacacgga cagggagagg gacacccacc tctggcagac ttctctggcg tgaggaatct
     1501 gtttggcgtg tgcgtgtaca gcttcatgtg ccagcactct ctgcccagcc tgatcacacc
     1561 cgtgagctct aagcggcacc tgaccagact ggtgtttctg gattacgtgc tgatcctggc
     1621 cttctatggc ctgctgagct ttacagccat cttctgcttt cgcggcgatt ccctgatgga
     1681 catgtatacc ctgaacttcg caaggtgtga cgtggtggga ctggcagccg tgcgcttctt
     1741 tctgggcctg tttccagtgt tcacaatctc taccaatttt cccatcatcg ccgtgacact
     1801 gcggaacaat tggaagaccc tgttccacag agagggcggc acataccctt gggtggtgga
     1861 tagagtggtg tttccaacaa tcaccctggt gccacccgtg ctggtggcct tctgcaccca
     1921 cgacctggag tccctggtgg gaatcacagg agcatacgca ggaaccggaa tccagtatgt
     1981 gatcccagcc tttctggtgt atcactgccg gagagataca cagctggcct tcggctgtgg
     2041 cgtgtccaac aagcacaggt ctccctttcg ccacaccttc tgggtgggct ttgtgctgct
     2101 gtgggccttc tcctgtttca tctttgtgac agccaatatc atcctgtctg agaccaagct
     2161 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2221 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2281 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2341 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2401 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2461 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2521 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2581 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2641 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2701 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2761 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2821 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2881 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2941 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3001 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3061 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3121 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3181 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3241 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3301 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3721 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3781 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3841 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3901 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3961 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4021 gccttttgct cacatgtt
//
