LOCUS       sequence-262985-        3948 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 3948)
  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 3948)
  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-NIPAL4
FEATURES             Location/Qualifiers
     source          1..3948
                     /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(2073..2172)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2189..2208)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2190..2208)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2213..2229)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2213..2229)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2331..2350)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2342..3151
                     /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(2409..2428)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3298..3886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3787..3806
                     /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 accatgcctg gcgacagctc cccaggcacc ctgcctctgt gggatgcatc
      721 tctgagccca cctctgggac ctgacccagg aggcttctct agggcaagcc acgcaggcga
      781 taagtctaga ccaccagcac ctgagctggg aagcccagga gccgtgcggc ccagagtggg
      841 ctcctgcgcc cccggcccta tggagctgcg ggtgtccaac acctcttgtg agaatggctc
      901 tctgctgcac ctgtactgct ctagccagga ggtgctgtgc cagatcgtga acgacctgtc
      961 ccctgaggtg ccatctaatg ccacattcca cagctggcag gagaggatcc gccagaacta
     1021 cggcttttat atcggcctgg gcctggcctt cctgtcctct tttctgatcg gcagcagcgt
     1081 gatcctgaag aagaagggcc tgctgaggct ggtggcaacc ggagcaacaa gggcagtgga
     1141 cggcggcttc ggctacctga aggatgccat gtggtgggca ggctttctga ccatggcagc
     1201 aggagaggtg gcaaacttcg gagcctatgc ctttgcccct gcaaccgtcg tgacaccact
     1261 gggcgccctg tctgtgctga tcagcgccat cctgtctagc tatttcctga gagagagcct
     1321 gaatctgctg ggcaagctgg gatgcgtgat ctgcgtggca ggatccaccg tgatggtcat
     1381 ccacgcccca gaggaggaga aggtgaccac aatcatggag atggcctcta agatgaagga
     1441 tacaggcttc atcgtgtttg ccgtgctgct gctggtgagc tgcctgatcc tgatcttcgt
     1501 gatcgccccc cggtacggcc agagaaacat cctgatctat atcatcatct gtagcgtgat
     1561 cggagccttt tccgtggcag cagtgaaggg actgggaatc accatcaaga atttctttca
     1621 gggactgcca gtggtgaggc acccactgcc ctacatcctg agcctgatcc tggccctgtc
     1681 cctgtctacc caggtgaact tcctgaatcg cgccctggat atcttcaaca catccctggt
     1741 gtttcctatc tactacgtgt tcttcaccac agtggtggtg acatcctcta tcatcctgtt
     1801 caaggagtgg tatagcatgt ccgccgtgga catcgcaggc accctgagcg gctttgtgac
     1861 aatcatcctg ggcgtgttca tgctgcacgc ctttaaggac ctggatatca gctgcgcctc
     1921 cctgccccac atgcacaaga atcctccacc atcccctgca ccagagccaa ccgtgatcag
     1981 gctggaggac aagaacgtgc tggtggataa tatcgagctg gcctccacaa gctccccaga
     2041 ggagaagccc aaggtgttta tcatccactc tgacccagct ttcttgtaca aagttggcat
     2101 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2161 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2221 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2281 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2341 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2401 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2461 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2521 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2581 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2641 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2701 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2761 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2821 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2881 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2941 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3001 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3061 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3121 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3181 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3241 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3301 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3361 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3421 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3481 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3541 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3601 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3661 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3721 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3781 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3841 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3901 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
