LOCUS       sequence-436381-        3783 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 3783)
  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 3783)
  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-NIPAL1_STOP
FEATURES             Location/Qualifiers
     source          1..3783
                     /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(1908..2007)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2024..2043)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2025..2043)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2048..2064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2048..2064)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2166..2185)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2177..2986
                     /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(2244..2263)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3133..3721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3622..3641
                     /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 accatgggag cacaggtgcg gctgccacct ggagagccct gcagagaggg
      721 ctacgtgctg tccctggtgt gccctaacag ctcccaggcc tggtgtgaga tcaccaatgt
      781 gagccagctg ctggcatccc ccgtgctgta cacagacctg aactattcca tcaacaatct
      841 gtctatcagc gccaacgtgg agaataagta ctctctgtat gtgggcctgg tgctggccgt
      901 gtctagctcc atcttcatcg gctctagctt catcctgaag aagaagggcc tgctgcagct
      961 ggccagcaag ggcttcacca gggcaggaca gggaggacac tcctacctga aggagtggct
     1021 gtggtgggtg ggactgctga gcatgggagc aggagaggca gcaaatttcg cagcctatgc
     1081 ctttgcccct gccaccctgg tgacaccact gggcgccctg agcgtgctga tctccgccat
     1141 cctgtcctct tactttctga acgagcacct gaatatccac ggcaagatcg gctgcatcct
     1201 gtctatcctg ggcagcaccg tgatggtcat ccacgcccca caggaggagg aggtgacatc
     1261 cctgcacgag atggagatga agctgcgcga ccccggcttc atctcctttg ccgtgatcat
     1321 caccgtgatc tctctggtgc tgatcctgat cgtggcccct aagaagggcc agacaaacat
     1381 cctggtgtat atctccatct gttctctgat cggcgccttc agcgtgagct ccgtgaaggg
     1441 cctgggcatc gccatcaagg agctgatcga gtggaagcca gtgtacaagc accccctggt
     1501 gttcgtgctg ctggccgtgc tggtgctgag cgtgaccaca cagatcaact atctgaataa
     1561 ggccctggat acctttaaca catccctggt gaccccaatc tactacgtgt tcttcacctc
     1621 tatggtggtg acatgcagcg ccatcctgtt tcaggagtgg tacggcatga ccgccggcga
     1681 catcatcggc accctgtctg gcttctttac aatcatcatc ggcatcttcc tgctgcacgc
     1741 ctttaagaat accgatatca catggtccga gctgacctct acagccaaga aggaggccgt
     1801 gagcctgaac gtgaatgaga acaattacgt gctgctggag aacctggagt gttctgcccc
     1861 tggctataat gacgatgtga ccctgttcag ccggacagac gattgagacc cagctttctt
     1921 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     1981 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2041 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2101 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2161 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2221 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2281 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2341 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2401 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2461 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2521 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2581 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2641 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2701 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2761 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2821 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2881 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2941 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3001 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3061 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3781 gtt
//
