LOCUS       sequence-261919-        3933 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 3933)
  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 3933)
  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_SLC46A3
FEATURES             Location/Qualifiers
     source          1..3933
                     /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(2058..2157)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2174..2193)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2175..2193)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2198..2214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2316..2335)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2327..3136
                     /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(2394..2413)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3283..3871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3772..3791
                     /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 accatgaaga tcctgttcgt ggagcctgcc atctttctga gcgccttcgc
      721 catgaccctg acaggcccac tgaccacaca gtacgtgtat cggagaatct gggaggagac
      781 cggcaattac accttcagct ccgacagcaa catctccgag tgtgagaaga ataagtctag
      841 ccctatcttc gcctttcagg aggaggtgca gaagaaggtg tcccggttca acctgcagat
      901 ggacatctcc ggcctgatcc caggcctggt gtctaccttt atcctgctga gcatctccga
      961 tcactacggc agaaagttcc ccatgatcct gtcctctgtg ggcgccctgg ccaccagcgt
     1021 gtggctgtgc ctgctgtgct atttcgcctt tccattccag ctgctgatcg ccagcacctt
     1081 catcggcgcc ttctgcggca attacaccac attttggggc gcctgcttcg cctatatcgt
     1141 ggaccagtgt aaggagcaca agcagaagac catcaggatc gccatcatcg attttctgct
     1201 gggactggtg accggactga caggactgag ctccggctac ttcatccgcg agctgggctt
     1261 tgagtggagc ttcctgatca tcgccgtgtc cctggccgtg aacctgatct atatcctgtt
     1321 ctttctgggc gatcccgtga aggagtgctc tagccagaac gtgaccatgt cttgtagcga
     1381 gggctttaag aatctgttct accggacata tatgctgttc aagaacgcca gcggcaagag
     1441 gcgctttctg ctgtgcctgc tgctgttcac cgtgatcaca tacttctttg tggtcatcgg
     1501 catcgcccca atctttatcc tgtatgagct ggacagcccc ctgtgctgga atgaggtgtt
     1561 catcggatac ggatccgccc tgggatccgc ctcttttctg accagcttcc tgggcatctg
     1621 gctgttctcc tattgcatgg aggatatcca catggccttt atcggcatct tcaccacaat
     1681 gaccggcatg gccatgacag cctttgcctc caccacactg atgatgttcc tggccagggt
     1741 gccctttctg ttcaccatcg tgcctttttc cgtgctgcgg tctatgctga gcaaggtggt
     1801 gagatctacc gagcagggca cactgtttgc ctgtatcgcc ttcctggaga cactgggagg
     1861 agtgaccgcc gtgtctacat tcaatggcat ctacagcgcc accgtggcct ggtatcccgg
     1921 ctttacattc ctgctgtctg ccggactgct gctgctgcct gcaatcagcc tgtgcgtggt
     1981 gaagtgtacc tcctggaacg agggctctta cgagctgctg atccaggagg agtcctctga
     2041 ggacgccagc gatcgcgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2101 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2161 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2221 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2281 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2341 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2401 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2461 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2521 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2581 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2641 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2701 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2761 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2821 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2881 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2941 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3001 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3061 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3121 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3181 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3241 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3301 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3361 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3421 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3481 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3541 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3601 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3661 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3721 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3781 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3841 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3901 cctggccttt tgctggcctt ttgctcacat gtt
//
