LOCUS       sequence-261147-        4257 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 4257)
  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 4257)
  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_SLC43A2
FEATURES             Location/Qualifiers
     source          1..4257
                     /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(2382..2481)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2498..2517)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2499..2517)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2522..2538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2522..2538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2640..2659)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2651..3460
                     /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(2718..2737)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3607..4195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4096..4115
                     /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 accatggcac ccaccctggc aacagcacac cggagaaggt ggtggatggc
      721 atgcaccgcc gtgctggaga acctgctgtt ctctgccgtg ctgctgggat ggggaagcct
      781 gctgatcatg ctgaagagcg agggctttta ctcctatctg tgcaccgagc cagagaacgt
      841 gaccaatgga acagtgggag gaacagcaga gccaggacac gaggaggtgt cctggatgaa
      901 cggctggctg tcttgtcagg cccaggacga gatgctgaat ctggccttca ccgtgggctc
      961 ctttctgctg tctgccatca cactgcccct gggcatcgtg atggataagt acggccctag
     1021 aaagctgagg ctgctgggat ccgcctgctt cgccgtgtct tgtctgctga tcgcctatgg
     1081 cgccagcaag cctaacgccc tgtccgtgct gatctttatc gccctggccc tgaatggctt
     1141 cggcggcatg tgcatgacct ttacaagcct gaccctgcca aatatgttcg gcgacctgcg
     1201 gagcaccttc atcgccctga tgatcggctc ttacgcaagc tccgccgtga ccttcccagg
     1261 catcaagctg atctatgatg ccggcgtgtc ttttatcgtg gtgctggtgg tgtgggcagg
     1321 atgcagcggc ctggtgttcc tgaactgttt ctttaattgg ccactggagc cctttcctgg
     1381 cccagaggac atggattact ctgtgaagat caagttcagc tggctgggct ttgaccacaa
     1441 gatcaccggc aagcagttct ataagcaggt gaccacagtg ggccgccggc tgagcgtggg
     1501 ctctagcatg cggtccgcca aggagcaggt ggccctgcag gagggccaca agctgtgcct
     1561 gagcacagtg gacctggagg tgaagtgtca gcctgatgcc gccgtggccc catccttcat
     1621 gcacagcgtg ttttccccca tcctgctgct gtctctggtg accatgtgcg tgacacagct
     1681 gcgcctgatc ttctacatgg gcgccatgaa caatatcctg aagtttctgg tgagcggcga
     1741 tcagaagacc gtgggcctgt acacatccat cttcggcgtg ctgcagctgc tgtgcctgct
     1801 gaccgcacct gtgatcggct atatcatgga ctggcggctg aaggagtgtg aggatgcctc
     1861 cgaggagcca gaggagaagg acgccaacca gggcgagaag aagaagaaga agcgggatcg
     1921 gcagatccag aagatcacca acgccatgag agccttcgcc tttacaaatc tgctgctggt
     1981 gggcttcggc gtgacctgtc tgatccccaa tctgcctctg cagatcctga gcttcatcct
     2041 gcacacaatc gtgaggggct ttatccactc cgccgtggga ggactgtacg cagccgtgta
     2101 tccatctacc cagtttggca gcctgacagg cctgcagtct ctgatcagcg ccctgttcgc
     2161 actgctgcag cagcctctgt ttctggcaat gatgggacca ctgcagggcg acccactgtg
     2221 ggtgaacgtg ggactgctgc tgctgtccct gctgggcttc tgcctgcctc tgtacctgat
     2281 ctgttataga aggcagctgg agaggcagct gcagcagagg caggaggacg ataagctgtt
     2341 cctgaagatc aacggctcct ctaatcagga ggcctttgtg gacccagctt tcttgtacaa
     2401 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2461 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2521 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2581 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2641 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2701 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2761 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2821 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2881 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2941 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3001 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3061 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3121 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3181 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3241 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3301 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3361 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3421 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3481 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3541 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3601 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3661 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3721 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3781 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3841 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3961 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4081 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4141 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4201 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
