LOCUS       sequence-262981-        3630 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 3630)
  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 3630)
  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
FEATURES             Location/Qualifiers
     source          1..3630
                     /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(1755..1854)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1871..1890)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1872..1890)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1895..1911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1895..1911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2013..2032)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2024..2833
                     /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(2091..2110)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2980..3568
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3469..3488
                     /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 ttcgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1801 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1861 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1921 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1981 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2041 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2101 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2161 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2221 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2281 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2341 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2401 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2461 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2521 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2581 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2641 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2701 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2761 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2821 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2881 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3601 ggccttttgc tggccttttg ctcacatgtt
//
