LOCUS       sequence-260407-        4608 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 4608)
  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 4608)
  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_SLC22A23
FEATURES             Location/Qualifiers
     source          1..4608
                     /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"
     regulatory      1626..1635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2733..2832)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2849..2868)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2850..2868)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2873..2889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2873..2889)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2991..3010)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3002..3811
                     /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(3069..3088)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3958..4546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4447..4466
                     /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 accatggcca tcgatcggag aagggaggca gcaggaggag gaccaggccg
      721 ccagcctgca ccagcagagg agaacggatc tctgccacct ggcgacgccg ccgccagcgc
      781 cccactggga ggaagggcag gcccaggagg aggagccgag atccagccac tgccaccact
      841 gcaccctgga ggaggccccc acccttcttg ctgtagcgcc gcagcagcac cttccctgct
      901 gctgctggac tacgatggct ctgtgctgcc attcctggga ggactgggag gaggatatca
      961 gaagacactg gtgctgctga cctggatccc cgccctgttc atcggcttta gccagttctc
     1021 tgacagcttc ctgctggatc agcctaactt ttggtgcagg ggagcaggca agggaaccga
     1081 gctggcagga gtgaccacaa ccggaagggg aggcgatatg ggcaattgga caagcctgcc
     1141 aacaaccccc ttcgccaccg caccatggga ggcagcagga aaccgctcca atagctccgg
     1201 agcagacgga ggcgataccc ctccactgcc aagcccacct gacaagggcg ataacgcctc
     1261 caattgcgac tgtagagcct gggattacgg catcagggcc ggcctggtgc agaacgtggt
     1321 gtctaagtgg gacctggtgt gcgataatgc ctggaaggtg cacatcgcca agttcagcct
     1381 gctggtgggc ctgatctttg gctatctgat cacaggctgt atcgcagact gggtgggccg
     1441 ccggcctgtg ctgctgtttt ctatcatctt catcctgatc tttggcctga ccgtggccct
     1501 gagcgtgaac gtgacaatgt tctccaccct gagattcttt gagggctttt gcctggccgg
     1561 catcatcctg acactgtacg cactgaggat cgagctgtgc ccaccaggca agcggttcat
     1621 gatcaccatg gtggcatcct tcgtggcaat ggcaggacag tttctgatgc caggactggc
     1681 cgccctgtgc agggattggc aggtgctgca ggccctgatc atctgtccat tcctgctgat
     1741 gctgctgtat tggagcatct ttcccgagtc cctgcggtgg ctgatggcca cacagcagtt
     1801 cgagagcgcc aagagactga tcctgcactt tacccagaag aacaggatga atcccgaggg
     1861 cgacatcaag ggcgtgatcc ctgagctgga gaaggagctg tccagaaggc caaagaaggt
     1921 gtgcatcgtg aaggtggtgg gcacaagaaa cctgtggaag aatatcgtgg tgctgtgcgt
     1981 gaactccctg accggctacg gcatccacca ctgtttcgcc aggtctatga tgggccacga
     2041 ggtgaaggtg cccctgctgg agaacttcta cgccgattac tatacaaccg cctctatcgc
     2101 cctggtgagc tgcctggcca tgtgcgtggt ggtgagattc ctgggccgga gaggaggact
     2161 gctgctgttt atgatcctga ccgccctggc aagcctgctg cagctgggac tgctgaatct
     2221 gatcggcaag tactcccagc accctgactc tggcatgtcc gactctgtga aggataagtt
     2281 ctccatcgcc ttttctatcg tgggaatgtt cgcatcccac gcagtgggaa gcctgtccgt
     2341 gttcttttgc gccgagatca cacctaccgt gatcagatgt ggaggactgg gactggtgct
     2401 ggcatctgcc ggatttggca tgctgacagc cccaatcatc gagctgcaca accagaaggg
     2461 ctatttcctg caccacatca tctttgcctg ctgtaccctg atctgcatca tctgtatcct
     2521 gctgctgcct gagtctagag atcagaatct gccagagaac atcagcaatg gcgagcacta
     2581 cacaaggcag cctctgctgc cacacaagaa gggagagcag ccactgctgc tgaccaacgc
     2641 cgagctgaag gactattccg gactgcacga tgcagcagca gcaggcgaca cactgcctga
     2701 gggcgccacc gccaatggca tgaaggccat ggacccagct ttcttgtaca aagttggcat
     2761 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2821 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2881 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2941 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     3001 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3061 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3121 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3181 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3241 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3301 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3361 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3421 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3481 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3541 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3601 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3661 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3721 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3781 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3841 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3901 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3961 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4021 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4081 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4141 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4201 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4261 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4321 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4381 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4441 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4501 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4561 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
