LOCUS       sequence-444349-        3987 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 3987)
  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 3987)
  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-DIRC2_STOP
FEATURES             Location/Qualifiers
     source          1..3987
                     /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(2112..2211)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2228..2247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2229..2247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2252..2268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2252..2268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2370..2389)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2381..3190
                     /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(2448..2467)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3337..3925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3826..3845
                     /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 accatgggat ccaggtggag ctccgaggag gagcgccagc ccctgctggg
      721 ccctggcctg ggcccaggcc tgggcgcctc ctggaggtct agggaggcag cagcagccgc
      781 cctgcctgca gccgtgcccg gccctggcag ggtgtacggc cggagatggc tggtgctgct
      841 gctgttctcc ctgctggcct ttgtgcaggg actggtgtgg aacacctggg gaccaatcca
      901 gaacagtgct cgacaagctt atggattttc tagctgggac atcgccctgc tggtgctgtg
      961 gggaccaatc ggcttcctgc cttgcttcgc ctttatgtgg ctgctggata agcggggcct
     1021 gagaatcacc gtgctgctga caagctttct gatggtgctg ggaaccggac tgaggtgtat
     1081 ccctatctcc gacctgatcc tgaagaggcg cctgatccac ggaggacaga tgctgaacgg
     1141 actggcaggc ccaacagtga tgaatgccgc ccccttcctg tccaccacat ggttttctgc
     1201 cgatgagaga gccaccgcca cagccatcgc aagcatgctg tcctacctgg gaggagcatg
     1261 cgccttcctg gtgggacctc tggtggtgcc agcaccaaac ggcacctccc cactgctggc
     1321 agcagagtcc tctagggccc acatcaagga cagaatcgag gccgtgctgt acgccgagtt
     1381 cggcgtggtg tgcctgatct tttctgccac actggcatat ttcccaccta ggccaccact
     1441 gcctccaagc gtggcagcag catctcagag actgagctac cggcggagcg tgtgcaggct
     1501 gctgtctaat ttccgctttc tgatgatcgc cctggcatat gcaatcccac tgggcgtgtt
     1561 tgcaggatgg agcggcgtgc tggacctgat cctgacccct gcacacgtga gccaggtgga
     1621 tgcaggatgg atcggcttct ggtccatcgt gggaggatgc gtggtgggaa tcgcaatggc
     1681 aaggttcgcc gacttcatca gaggcatgct gaagctgatc ctgctgctgc tgtttagcgg
     1741 cgccaccctg agctccacat ggttcaccct gacatgcctg aactccatca cccacctgcc
     1801 actgaccaca gtgacactgt acgcctcttg tatcctgctg ggcgtgtttc tgaattctag
     1861 cgtgcccatc ttctttgagc tgttcgtgga gaccgtgtat cctgtgccag agggcatcac
     1921 ctgcggcgtg gtgacattcc tgagcaacat gtttatgggc gtgctgctgt tctttctgac
     1981 cttttatcac acagagctgt cctggttcaa ttggtgcctg cccggctctt gtctgctgag
     2041 cctgctgctg atcctgtgct tcagggagtc ttacgaccgc ctgtatctgg atgtggtggt
     2101 gagcgtgtaa gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2161 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2221 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2341 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2401 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2461 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2521 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2581 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2641 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2701 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2761 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2821 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2881 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2941 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3001 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3061 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3121 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3181 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3241 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3481 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3961 cttttgctgg ccttttgctc acatgtt
//
