LOCUS       sequence-261086-        4227 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 4227)
  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 4227)
  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_SLC43A1
FEATURES             Location/Qualifiers
     source          1..4227
                     /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(2352..2451)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2468..2487)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2469..2487)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2492..2508)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2492..2508)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2610..2629)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2621..3430
                     /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(2688..2707)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3577..4165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4066..4085
                     /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 accatggcac ctaccctgca gcaggcatac cggagaaggt ggtggatggc
      721 ctgtacagcc gtgctggaga acctgttctt ttccgccgtg ctgctgggat ggggatctct
      781 gctgatcatc ctgaagaacg agggcttcta tagctccacc tgccctgccg agtctagcac
      841 aaataccaca caggacgagc agcgccggtg gccaggctgt gaccagcagg atgagatgct
      901 gaatctgggc ttcaccatcg gctcttttgt gctgagcgcc accacactgc cactgggcat
      961 cctgatggac agattcggac caagacctgt gaggctggtg ggatccgcct gctttaccgc
     1021 ctcttgtaca ctgatggcac tggcaagcag ggatgtggag gccctgtccc ctctgatctt
     1081 cctggccctg tccctgaacg gattcggagg catctgcctg acctttacat ctctgaccct
     1141 gccaaacatg ttcggcaatc tgcggtctac actgatggcc ctgatgatcg gcagctacgc
     1201 ctcctctgcc atcacctttc caggcatcaa gctgatctat gacgccggcg tggccttcgt
     1261 ggtcatcatg tttacctgga gcggcctggc ctgcctgatc ttcctgaact gtacactgaa
     1321 ttggccaatc gaggcatttc cagcaccaga ggaggtgaac tacaccaaga agatcaagct
     1381 gtccggactg gccctggacc acaaggtgac cggcgatctg ttctatacac acgtgaccac
     1441 aatgggacag cgcctgagcc agaaggcacc atccctggag gacggctctg atgcctttat
     1501 gagcccccag gatgtgcggg gcacctctga gaatctgcct gagcgcagcg tgccactgcg
     1561 gaagtccctg tgctctccca cattcctgtg gagcctgctg accatgggca tgacacagct
     1621 gcgcatcatc ttttacatgg ccgccgtgaa caagatgctg gagtatctgg tgaccggcgg
     1681 ccaggagcac gagacaaatg agcagcagca gaaggtggcc gagaccgtgg gcttctacag
     1741 ctccgtgttt ggcgccatgc agctgctgtg cctgctgaca tgtcccctga tcggctatat
     1801 catggattgg agaatcaagg actgcgtgga tgcacctacc cagggaacag tgctgggcga
     1861 cgcaagggat ggagtggcaa ccaagagcat cagacctagg tactgcaaga tccagaagct
     1921 gacaaacgcc atctccgcct tcaccctgac aaatctgctg ctggtgggct ttggcatcac
     1981 ctgtctgatc aacaatctgc acctgcagtt cgtgaccttt gtgctgcaca caatcgtgcg
     2041 gggcttcttt cactccgcct gcggatctct gtacgcagcc gtgttcccct ccaatcactt
     2101 tggcaccctg acaggcctgc agagcctgat ctccgccgtg ttcgcactgc tgcagcagcc
     2161 tctgtttatg gcaatggtgg gacccctgaa gggagagcct ttctgggtga acctgggcct
     2221 gctgctgttc agcctgctgg gctttctgct gccatcctac ctgttttact atagggcaag
     2281 gctgcagcag gagtatgcag caaatggcat gggccccctg aaggtgctgt ctggcagcga
     2341 ggtgaccgcc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2401 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2461 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     2521 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2581 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2641 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2701 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2761 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2821 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2881 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2941 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3001 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3061 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3121 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3181 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3241 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3301 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3361 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3421 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3481 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3721 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4201 cttttgctgg ccttttgctc acatgtt
//
