LOCUS       sequence-261148-        3927 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 3927)
  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 3927)
  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_SLC46A1
FEATURES             Location/Qualifiers
     source          1..3927
                     /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(2052..2151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2168..2187)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2169..2187)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2192..2208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2192..2208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2310..2329)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2321..3130
                     /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(2388..2407)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3277..3865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3766..3785
                     /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 accatggagg gatctgccag cccacctgag aagccaaggg caaggccagc
      721 agcagccgtg ctgtgcaggg gccctgtgga gccactggtg ttcctggcca actttgccct
      781 ggtgctgcag ggcccactga ccacacagta cctgtggcac cggttcagcg ccgacctggg
      841 atataacgga accaggcaga gaggcggctg tagcaataga tccgccgatc ccaccatgca
      901 ggaggtggag accctgacat cccactggac actgtacatg aatgtgggcg gcttcctggt
      961 gggcctgttt agctccacac tgctgggagc atggtccgac tctgtgggcc ggagacctct
     1021 gctggtgctg gcaagcctgg gactgctgct gcaggccctg gtgtccgtgt tcgtggtgca
     1081 gctgcagctg cacgtgggct attttgtgct gggcagaatc ctgtgcgcac tgctgggcga
     1141 cttcggagga ctgctggcag catcttttgc cagcgtggca gacgtgagca gctccaggag
     1201 ccgcaccttc cggatggcac tgctggaggc atccatcgga gtggcaggaa tgctggcatc
     1261 tctgctggga ggacactggc tgagagcaca gggctacgcc aacccctttt ggctggccct
     1321 ggccctgctg atcgccatga ccctgtatgc cgccttctgt tttggcgaga cactgaagga
     1381 gcctaagtct accaggctgt tcacatttcg gcaccacaga agcatcgtgc agctgtacgt
     1441 ggcaccagca ccagagaagt ctcgcaagca cctggccctg tatagcctgg ccatcttcgt
     1501 ggtcatcacc gtgcactttg gcgcccagga catcctgacc ctgtacgagc tgagcacacc
     1561 tctgtgctgg gattccaagc tgatcggata cggatctgcc gcacagcacc tgccatatct
     1621 gacaagcctg ctggccctga agctgctgca gtactgtctg gcagacgcat gggtggcaga
     1681 gatcggactg gccttcaaca tcctgggcat ggtggtgttc gcctttgcca ccatcacacc
     1741 actgatgttc accggctatg gcctgctgtt tctgtccctg gtcatcacac ccgtgatcag
     1801 ggccaagctg tctaagctgg tgcgcgagac cgagcagggc gccctgttta gcgccgtggc
     1861 atgcgtgaat tccctggcca tgctgaccgc ctccggcatc ttcaactctc tgtatcctgc
     1921 caccctgaac ttcatgaagg gcttcccatt tctgctggga gcaggactgc tgctgatccc
     1981 agccgtgctg atcggcatgc tggagaaggc cgatcctcac ctggagttcc agcagtttcc
     2041 ccagtcccct gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2101 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2161 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2221 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2281 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2341 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2401 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2461 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2521 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2581 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2641 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2701 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2761 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2821 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2881 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2941 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3001 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3061 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3121 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3181 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3421 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3901 cttttgctgg ccttttgctc acatgtt
//
