LOCUS       sequence-420957-        3633 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 3633)
  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 3633)
  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-NIPA2_STOP
FEATURES             Location/Qualifiers
     source          1..3633
                     /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(1758..1857)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1874..1893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1875..1893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1898..1914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1898..1914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2016..2035)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2027..2836
                     /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(2094..2113)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2983..3571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3472..3491
                     /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 accatgtccc agggcagggg caagtacgac ttctatatcg gcctgggcct
      721 ggccatgagc tcctctatct tcatcggcgg cagcttcatc ctgaagaaga agggactgct
      781 gaggctggca aggaagggat ccatgagggc aggacaggga ggacacgcct acctgaagga
      841 gtggctgtgg tgggcaggac tgctgagcat gggagcagga gaggtggcaa atttcgcagc
      901 ctatgccttt gccccagcca ccctggtgac accactgggc gccctgagcg tgctggtgtc
      961 cgccatcctg agctcctact ttctgaatga gcggctgaac ctgcacggca agatcggctg
     1021 cctgctgtct atcctgggca gcaccgtgat ggtcatccac gcccctaagg aggaggagat
     1081 cgagacactg aatgagatgt cccacaagct gggcgatcca ggcttcgtgg tgtttgccac
     1141 cctggtggtc atcgtggccc tgatcctgat cttcgtggtg ggaccaaggc acggacagac
     1201 caacatcctg gtgtacatca caatctgctc tgtgatcggc gccttttctg tgagctgcgt
     1261 gaagggcctg ggcatcgcca tcaaggagct gttcgcaggc aagcccgtgc tgaggcaccc
     1321 tctggcctgg atcctgctgc tgtctctgat cgtgtgcgtg agcacccaga tcaattatct
     1381 gaacagagcc ctggacatct ttaacacctc tatcgtgaca cccatctact acgtgttctt
     1441 caccacaagc gtgctgacat gttccgccat cctgttcaag gagtggcagg atatgcctgt
     1501 ggacgatgtg atcggcaccc tgagcggctt ctttacaatc atcgtgggca tcttcctgct
     1561 gcacgccttt aaggacgtgt ccttctctct ggcctccctg cccgtgtctt ttagaaagga
     1621 tgagaaggcc atgaatggca acctgtccaa tatgtatgag gtgctgaaca ataacgagga
     1681 gtctctgacc tgtggcatcg agcagcacac aggcgagaac gtgtcccgga gaaatggcaa
     1741 cctgaccgcc ttctgagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1801 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1861 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1921 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1981 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2041 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2101 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2161 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2221 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2281 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2341 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2401 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2461 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2521 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2581 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2641 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2701 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2761 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2821 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2881 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3121 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3601 cctggccttt tgctggcctt ttgctcacat gtt
//
