LOCUS       sequence-259254-        3885 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 3885)
  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 3885)
  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_SLC52A2
FEATURES             Location/Qualifiers
     source          1..3885
                     /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(2010..2109)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2126..2145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2127..2145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2150..2166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2150..2166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2268..2287)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2279..3088
                     /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(2346..2365)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3235..3823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3724..3743
                     /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 accatggcag cacctacccc agcaaggccc gtgctgacac acctgctggt
      721 ggccctgttc ggaatgggat cctgggcagc agtgaacgga atctgggtgg agctgcctgt
      781 ggtggtgaag gagctgccag agggatggag cctgccatcc tacgtgtctg tgctggtggc
      841 cctgggcaat ctgggactgc tggtggtgac cctgtggcgg agactggcac ctggcaagga
      901 tgagcaggtg ccaatcaggg tggtgcaggt gctgggaatg gtgggaacag ccctgctggc
      961 aagcctgtgg caccacgtgg caccagtggc aggacagctg cactccgtgg ccttcctggc
     1021 cctggccttt gtgctggccc tggcatgctg tgcctccaac gtgaccttcc tgccttttct
     1081 gtctcacctg ccccctaggt tcctgcgctc cttctttctg ggacagggac tgtctgccct
     1141 gctgccatgc gtgctggccc tggtgcaggg agtgggccgg ctggagtgtc caccagcacc
     1201 catcaatgga acccctggac ctccactgga tttcctggag agatttccag cctccacctt
     1261 cttttgggcc ctgacagccc tgctggtggc atctgccgca gcatttcagg gactgctgct
     1321 gctgctgccc cctccaccca gcgtgcccac aggcgagctg ggctccggac tgcaagtggg
     1381 agcaccagga gcagaggagg aggtggagga gagctcccca ctgcaggagc ctccatccca
     1441 ggcagcagga accacaccag gacctgaccc aaaggcctac cagctgctga gcgcccggtc
     1501 cgcctgcctg ctgggactgc tggccgccac caacgccctg acaaatggcg tgctgcccgc
     1561 cgtgcagtcc ttctcttgtc tgccttacgg caggctggcc tatcacctgg cagtggtgct
     1621 gggaagcgcc gcaaaccctc tggcatgctt cctggccatg ggcgtgctgt gccgctctct
     1681 ggcaggactg ggaggactga gcctgctggg cgtgttttgc ggaggatacc tgatggcact
     1741 ggccgtgctg agcccatgtc cacctctggt gggcacctcc gccggagtgg tgctggtggt
     1801 gctgtcttgg gtgctgtgcc tgggcgtgtt tagctatgtg aaggtggccg cctctagcct
     1861 gctgcacgga ggaggccggc ctgccctgct ggcagccggc gtggcaatcc aagtgggatc
     1921 tctgctggga gcagtggcaa tgttcccacc cacatctatc tatcacgtgt ttcacagcag
     1981 aaaggactgc gccgatccct gtgacagcga cccagctttc ttgtacaaag ttggcattat
     2041 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2101 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2161 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2221 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2281 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2341 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2401 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2461 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2521 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2581 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2641 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2701 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     2761 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2821 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2881 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2941 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3001 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3061 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3121 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3181 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3241 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3301 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3361 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3421 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3481 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3541 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3601 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3661 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3721 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3781 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3841 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
