LOCUS       sequence-354276-        3525 bp    DNA     circular SYN 15-DEC-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 3525)
  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 3525)
  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-TMEM165_STOP
FEATURES             Location/Qualifiers
     source          1..3525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1082..1181)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1198..1217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1199..1217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1222..1238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1222..1238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1340..1359)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1351..2160
                     /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(1418..1437)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2307..2895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2796..2815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3049..3066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3225..3252)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3344..3430)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3479..3501
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3493..3510
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3494..3510
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc agcagcagca
      121 cctggaaacg gaagggcatc tgccccaaga ctgctgctgc tgttcctggt gccactgctg
      181 tgggcaccag cagccgtgcg ggcaggacca gacgaggatc tgagccacag aaataaggag
      241 ccacctgcac cagcacagca gctgcagcca cagcccgtgg ccgtgcaggg ccctgagcca
      301 gcccgggtgg agaagatctt tacccccgcc gcccctgtgc acacaaacaa ggaggaccca
      361 gccacccaga caaatctggg cttcatccac gcctttgtgg ccgccatctc tgtgatcatc
      421 gtgagcgagc tgggcgataa gaccttcttt atcgccgcca tcatggccat gcggtacaac
      481 agactgaccg tgctggcagg agcaatgctg gccctgggcc tgatgacatg cctgtctgtg
      541 ctgttcggct acgccaccac agtgatccct cgggtgtata cctactatgt gagcacagtg
      601 ctgttcgcca tctttggcat caggatgctg cgcgagggcc tgaagatgtc cccagacgag
      661 ggacaggagg agctggagga ggtgcaggcc gagctgaaga agaaggacga ggagttccag
      721 agaaccaagc tgctgaatgg accaggcgat gtggagaccg gcacatccat cacagtgccc
      781 cagaagaagt ggctgcactt catctctcct atctttgtgc aggccctgac cctgacattt
      841 ctggcagagt ggggcgacag gtcccagctg accacaatcg tgctggcagc aagggaggat
      901 ccttatggag tggcagtggg aggaaccgtg ggacactgcc tgtgcacagg actggccgtg
      961 atcggaggaa ggatgatcgc ccagaagatc agcgtgcgca ccgtgacaat catcggcggc
     1021 atcgtgtttc tggccttcgc cttttccgcc ctgttcatca gccctgattc cggcttttga
     1081 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1141 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1201 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1261 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1321 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1381 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1441 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1501 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1561 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1621 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1681 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1741 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1801 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1861 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1921 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1981 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2041 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2101 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2161 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2221 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2281 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2341 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2401 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2461 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2521 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2581 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2641 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2701 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2761 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2821 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2881 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2941 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3001 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3061 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3121 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3181 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3241 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3301 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3361 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3421 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     3481 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
