LOCUS       sequence-260115-        4533 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 4533)
  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 4533)
  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_SLC24A2
FEATURES             Location/Qualifiers
     source          1..4533
                     /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(2658..2757)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2774..2793)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2775..2793)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2798..2814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2916..2935)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2927..3736
                     /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(2994..3013)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3883..4471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4372..4391
                     /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 accatggacc tgcagcagag caccacaatc acctccctgg agaagtggtg
      721 cctggatgag tctctgagcg gctgtcggag acactacagc gtgaagaaga agctgaagct
      781 gatcagggtg ctgggcctgt tcatgggcct ggtggccatc tccaccgtgt ccttttctat
      841 cagcgccttc tccgagaccg acacacagtc tacaggagag gcaagcgtgg tgtccggacc
      901 tagggtggca cagggatacc accagcgcac cctgctggac ctgaatgata agatcctgga
      961 ctatacacca cagccccctc tgtctaagga gggcgagtct gagaacagca ccgatcacgc
     1021 ccagggcgac taccccaagg atatcttttc cctggaggag aggcgcaagg gcgccatcat
     1081 cctgcacgtg atcggcatga tctatatgtt catcgccctg gccatcgtgt gcgatgagtt
     1141 ctttgtgcct tccctgaccg tgatcacaga gaagctgggc atctctgacg atgtggcagg
     1201 agccaccttt atggcagcag gaggcagcgc cccagagctg tttacatccc tgatcggcgt
     1261 gttcatcgcc cactctaatg tgggaatcgg aaccatcgtg ggaagcgccg tgttcaacat
     1321 cctgtttgtg atcggcatgt gcgccctgtt ctctcgggag atcctgaatc tgacatggtg
     1381 gcccctgttt agagacgtga gcttctacat cgtggatctg atcatgctga tcatcttctt
     1441 tctggacaat gtgatcatgt ggtgggagag cctgctgctg ctgaccgcct acttctgtta
     1501 tgtggtgttc atgaagttta acgtgcaggt ggagaagtgg gtgaagcaga tgatcaacag
     1561 gaataaggtg gtgaaggtga ccgcaccaga ggcacaggca aagccatccg ccgcaaggga
     1621 caaggatgag cctacactgc cagccaagcc aaggctgcag aggggaggaa gctccgcctc
     1681 cctgcacaat tctctgatgc ggaactccat ctttcagctg atgatccaca cactggaccc
     1741 actggccgag gagctgggct cttatggcaa gctgaagtac tatgatacca tgacagagga
     1801 gggccggttc agagagaagg ccagcatcct gcacaagatc gccaagaaga agtgtcacgt
     1861 ggatgagaac gagagacaga atggcgccgc caaccacgtg gagaagatcg agctgcccaa
     1921 ttccacctct acagacgtgg agatgacccc ctctagcgat gccagcgagc ctgtgcagaa
     1981 cggcaatctg tcccacaaca tcgagggagc agaggcacag acagcagacg aggaggagga
     2041 tcagcctctg tctctggcct ggccaagcga gacccggaag caggtgacat tcctgatcgt
     2101 gtttcctatc gtgttcccac tgtggatcac cctgcccgac gtgcggaagc cttcctctag
     2161 aaagttcttt cccatcacct tctttggctc catcacatgg atcgccgtgt tttcttatct
     2221 gatggtgtgg tgggcacacc aagtgggaga gaccatcggc atcagcgagg agatcatggg
     2281 actgaccatc ctggcagcag gcacatctat ccctgatctg atcacaagcg tgatcgtggc
     2341 aaggaaggga ctgggcgaca tggccgtgag ctcctctgtg ggctccaaca tcttcgatat
     2401 caccgtggga ctgccactgc catggctgct gtacacagtg atccaccgct ttcagcctgt
     2461 ggccgtgagc tccaatggcc tgttctgcgc catcgtgctg ctgtttatca tgctgctgtt
     2521 cgtgatcctg agcatcgccc tgtgcaagtg gcggatgaac aagatcctgg gcttcatcat
     2581 gtttggcctg tatttcgtgt ttctggtggt gagcgtgctg ctggaggaca gaatcctgac
     2641 ctgcccagtg tccatcgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2701 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2761 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2821 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2881 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2941 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     3001 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     3061 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     3121 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     3181 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     3241 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3301 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3361 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3421 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3481 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3541 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3601 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3661 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3721 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3781 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3841 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3901 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3961 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4021 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4081 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4141 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4201 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4261 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4321 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4381 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4441 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4501 cctggccttt tgctggcctt ttgctcacat gtt
//
