LOCUS       sequence-261197-        3975 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 3975)
  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 3975)
  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_SLC46A2
FEATURES             Location/Qualifiers
     source          1..3975
                     /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(2100..2199)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2216..2235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2217..2235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2240..2256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2240..2256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2358..2377)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2369..3178
                     /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(2436..2455)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3325..3913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3814..3833
                     /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 cagaggtgac atgcccaagg agaggacacc tgccaaggtt
      721 ccaccctaga acctgggtgg agcctgtggt ggcaagctcc caggtggcag caagcctgta
      781 cgacgcagga ctgctgctgg tggtgaaggc atcctatgga accggaggat ctagcaacca
      841 ctccgcctct ccaagcccaa ggggcgccct ggaggatcag cagcagcgcg ccatcagcaa
      901 cttctacatc atctataatc tggtggtggg actgagccca ctgctgtccg cctacggact
      961 gggatggctg tctgaccggt atcacagaaa gatctctatc tgcatgagcc tgctgggctt
     1021 tctgctgagc aggctgggcc tgctgctgaa ggtgctgctg gattggcctg tggaggtgct
     1081 gtacggagca gccgccctga atggactgtt cggcggcttt tccgccttct ggtctggcgt
     1141 gatggcactg ggatctctgg gatcctctga gggcaggcgc agcgtgcgcc tgatcctgat
     1201 cgacctgatg ctgggcctgg ccggcttttg tggcagcatg gcctccggcc acctgttcaa
     1261 gcagatggca ggacacagcg gacagggact gatcctgaca gcctgctccg tgtcttgtgc
     1321 ctctttcgcc ctgctgtaca gcctgctggt gctgaaggtg cctgagagcg tggccaagcc
     1381 ttcccaggag ctgccagcag tggacaccgt gtccggaacc gtgggaacat accggaccct
     1441 ggacccagat cagctggatc agcagtatgc agtgggacac ccaccttctc caggcaaggc
     1501 caagcctcac aagaccacaa tcgccctgct gtttgtgggc gccatcatct atgacctggc
     1561 cgtggtgggc acagtggatg tgatcccact gttcgtgctg agggagccac tgggatggaa
     1621 ccaggtgcaa gtgggatacg gaatggcagc aggctataca atctttatca cctccttcct
     1681 gggcgtgctg gtgttttcta ggtgcttccg cgataccaca atgatcatga tcggcatggt
     1741 gtcttttggc agcggcgccc tgctgctggc ctttgtgaag gagacctaca tgttctatat
     1801 cgcccgggcc gtgatgctgt tcgccctgat cccagtgacc acaatcagat ccgccatgtc
     1861 taagctgatc aagggcagct cctacggcaa ggtgtttgtg atcctgcagc tgtccctggc
     1921 cctgacaggc gtggtgacct ctacactgta caataagatc tatcagctga ccatggacat
     1981 gtttgtgggc tcctgtttcg ccctgtctag ctttctgagc ttcctggcca tcatccccat
     2041 ctccatcgtg gcctacaagc aggtgcctct gagcccatat ggcgatatca tcgagaagga
     2101 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2161 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2221 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2281 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2341 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2401 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2461 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2521 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2581 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2641 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2701 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2761 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2821 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2881 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2941 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3001 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3061 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3121 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3181 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3241 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3961 ttttgctcac atgtt
//
