LOCUS       sequence-262972-        3537 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 3537)
  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 3537)
  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-NIPA1
FEATURES             Location/Qualifiers
     source          1..3537
                     /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(1662..1761)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1778..1797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1779..1797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1802..1818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1802..1818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1920..1939)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1931..2740
                     /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(1998..2017)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2887..3475
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3376..3395
                     /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 accatgggca cagccgccgc agccgccgca gcagcagcag cagccgccgc
      721 cggcgagggc gcccggagcc cctcccctgc cgccgtgtct ctgggactgg gagtggcagt
      781 ggtgagctcc ctggtgaacg gcagcacatt cgtgctgcag aagaagggca tcgtgagggc
      841 caagcggaga ggcaccagct acctgacaga catcgtgtgg tgggcaggaa ccatcgcaat
      901 ggcagtggga cagatcggca acttcctggc ctataccgcc gtgcctacag tgctggtgac
      961 cccactgggc gccctgggag tgccttttgg cagcatcctg gcctcctacc tgctgaagga
     1021 gaagctgaat atcctgggca agctgggctg cctgctgtct tgtgccggca gcgtggtgct
     1081 gatcatccac tccccaaagt ctgagagcgt gaccacacag gccgagctgg aggagaagct
     1141 gacaaacccc gtgttcgtgg gctacctgtg catcgtgctg ctgatgctgc tgctgctgat
     1201 cttttggatc gccccagccc acggccccac caatatcatg gtgtatatct ccatctgttc
     1261 tctgctgggc tccttcacag tgccatctac caagggaatc ggactggcag cacaggacat
     1321 cctgcacaac aatccttcta gccagagggc cctgtgcctg tgcctggtgc tgctggccgt
     1381 gctgggatgc agcatcatcg tgcagtttcg ctatatcaac aaggccctgg agtgtttcga
     1441 ctccagcgtg ttcggcgcca tctactatgt ggtgttcacc acactggtgc tgctggccag
     1501 cgccatcctg tttcgggagt ggtccaatgt gggcctggtg gatttcctgg gcatggcctg
     1561 cggctttacc acagtgtccg tgggcatcgt gctgatccag gtgtttaagg agttcaactt
     1621 taatctgggc gagatgaaca agtctaatat gaagaccgat gacccagctt tcttgtacaa
     1681 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1741 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     1801 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     1861 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     1921 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     1981 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2041 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2101 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2161 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2221 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2281 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2341 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2401 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     2461 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     2521 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     2581 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     2641 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     2701 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     2761 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     2821 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2881 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2941 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3001 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3061 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3121 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3181 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3241 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3301 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3361 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3421 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3481 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
