LOCUS       sequence-261844-        5847 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 5847)
  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 5847)
  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_SLC24A1
FEATURES             Location/Qualifiers
     source          1..5847
                     /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(3972..4071)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4088..4107)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4089..4107)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4112..4128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4112..4128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4230..4249)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4241..5050
                     /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(4308..4327)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5197..5785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5686..5705
                     /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 agctgatccg gatgggcccc caggagagat ggctgctgag
      721 gacaaagcgc ctgcactggt ccagactgct gttcctgctg ggcatgctga tcatcggctc
      781 tacctatcag cacctgcgga gacccagagg actgagctcc ctgtgggcag ccgtgtctag
      841 ccaccagcct atcaagctgg cctctaggga cctgtcctct gaggagatga tgatgatgag
      901 ctcctctcct agcaagccaa gctccgagat gggcggcaag atgctggtgc ctcaggcaag
      961 cgtgggatcc gatgaggcca ccctgtccat gacagtggag aacatcccct ctatgcctaa
     1021 gaggaccgcc aagatgatcc ctaccacaac caagaacaat tactccccaa ccgcagcagg
     1081 aacagagagg aggaaggagg acacaccaac ctctagccgg acactgacct actatacaag
     1141 cacctcctct agacagatcg tgaagaagta cacacccacc cctaggggcg agatgaagtc
     1201 ttatagcccc acccaggtga gggagaaggt gaagtacaca ccatccccaa ggggccggag
     1261 agtgggaaca tatgtgccta gcacattcat gaccatggag acatcccacg ccatcacccc
     1321 acgcacaacc gtgaaggaca gcgatatcac agccacctac aagatcctgg agaccaactc
     1381 cctgaagcgg atcatggagg agacaacccc cacaaccctg aagggcatgt tcgacagcac
     1441 acctaccttt ctgacccacg aggtggaggc caatgtgctg acatctccaa gaagcgtgat
     1501 ggagaagaac aatctgttcc cccctaggcg cgtggagtct aacagctccg cccacccatg
     1561 gggactggtg ggcaagagca atccaaagac cccacaggga acagtgctgc tgcacacccc
     1621 tgccacatct gagggccagg tgaccatcag cacaatgacc ggctctagcc cagccgagac
     1681 caaggccttt acagccgcct ggtccctgag gaacccttct ccacgcacct ccgtgtctgc
     1741 catcaagaca gcaccagcaa tcgtgtggag gctggccaag aagcctagca ccgcaccaag
     1801 cacatccaca acccctaccg tgcgggccaa gctgacaatg caggtgcacc actgcgtggt
     1861 ggtgaagccc acccctgcca tgctgacaac cccatctccc agcctgacaa ccgccctgct
     1921 gccagaggag ctgtcccctt ctccaagcgt gctgccacca tccctgccag acctgcaccc
     1981 taagggagag tatcctccag atctgttctc tgtggaggag cggagacagg gctgggtggt
     2041 gctgcacgtg ttcggcatga tgtacgtgtt cgtggccctg gccatcgtgt gcgacgagta
     2101 cttcgtgcct gccctgggcg tgatcaccga caagctgcag atcagcgagg atgtggcagg
     2161 agccaccttc atggcagcag gaggctccgc cccagagctg ttcacctctc tgatcggcgt
     2221 gtttatctcc cactctaacg tgggaatcgg aaccatcgtg ggatccgccg tgttcaatat
     2281 cctgtttgtg atcggcacct gttccctgtt ttctcgcgag atcctgaacc tgacatggtg
     2341 gcccctgttc cgggacgtga gcttttatat cctggatctg atcatgctga tcctgttctt
     2401 tctggatagc ctgatcgcct ggtgggagtc cctgctgctg ctgctggcct acgccttcta
     2461 cgtgttcacc atgaaatgga ataagcacat cgaagtgtgg gtgaaggagc agctgtccag
     2521 gcgcccagtg gccaaagtga tggccctgga ggacctgtct aagccaggcg atggagcaat
     2581 cgcagtggac gagctgcagg ataacaagaa gctgaagctg ccctccctgc tgaccagagg
     2641 ctcctctagc acaagcctgc acaattccac catcaggtct acaatctacc agctgatgct
     2701 gcacagcctg gaccctctgc gggaggtgag actggccaag gagaaagagg aggagagcct
     2761 gaaccaggga gcaagggcac agccacaggc aaaggcagag tccaagccag aggaggagga
     2821 gccagcaaag ctgccagcag tgacagtgac cccagcacct gtgccagaca tcaagggcga
     2881 tcagaaggag aatccaggag gacaggagga tgtggcagag gcagagtcta caggagagat
     2941 gccaggagag gagggagaga ccgcaggaga gggcgagaca gaggagaaga gcggaggaga
     3001 gacccagcct gagggagagg gagagacaga gacccagggc aagggagagg agtgcgagga
     3061 cgagaacgag gccgagggca agggcgacaa tgagggcgag gatgagggcg agatccacgc
     3121 cgaggatggc gagatgaagg gcaacgaggg agagaccgag agccaggagc tgtccgccga
     3181 gaaccacgga gaggccaaga atgacgagaa gggagtggag gatggaggag gaagcgacgg
     3241 aggcgattcc gaggaagaag aggaggagga ggaggagcag gaagaggaag aagaagaaga
     3301 ggagcaggaa gaggaggaag aagaagagga agaagaagag gaaaagggca atgaggagcc
     3361 actgtccctg gactggcctg agacccgcca gaagcaggcc atctatctgt tcctgctgcc
     3421 tatcgtgttt ccactgtggc tgacagtgcc tgatgtgcgg agacaggaga gccggaagtt
     3481 cttcgtgttc accttcctgg gctctatcat gtggatcgcc atgttcagct acctgatggt
     3541 gtggtgggca caccaagtgg gagagacaat cggcatcagc gaggagatca tgggactgac
     3601 catcctggca gcaggcacat ctatcccaga cctgatcacc agcgtgatcg tggcaaggaa
     3661 gggactgggc gacatggccg tgtcctctag cgtgggcagc aacatcttcg atatcaccgt
     3721 gggactgcca gtgccttggc tgctgttttc cctgatcaac ggcctgcagc cagtgcccgt
     3781 gtcctctaat ggcctgtttt gcgccatcgt gctgctgttc ctgatgctgc tgtttgtgat
     3841 cagctccatc gcctcctgta agtggagaat gaataagatc ctgggcttca ccatgtttct
     3901 gctgtacttc gtgtttctga tcatctctgt gatgctggag gataggatca tctcttgtcc
     3961 cgtgagcgtg gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     4021 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     4081 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     4141 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4201 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4261 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     4321 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     4381 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     4441 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     4501 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     4561 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     4621 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     4681 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     4741 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     4801 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     4861 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     4921 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     4981 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     5041 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     5101 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5581 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5641 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5701 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5761 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5821 cttttgctgg ccttttgctc acatgtt
//
