LOCUS       sequence-416388-        4458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4458)
  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 4458)
  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-SLC7A4_STOP
FEATURES             Location/Qualifiers
     source          1..4458
                     /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(2583..2682)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2699..2718)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2700..2718)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2723..2739)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2723..2739)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2841..2860)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2852..3661
                     /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(2919..2938)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3808..4396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4297..4316
                     /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 accatggcaa ggggactgcc aacaatcgca tccctggcaa ggctgtgcca
      721 gaagctgaac agactgaagc ccctggagga cagcaccatg gagacatccc tgcggagatg
      781 tctgagcacc ctggatctga cactgctggg agtgggagga atggtgggat ccggactgta
      841 cgtgctgacc ggagcagtgg caaaggaggt ggcaggacct gccgtgctgc tgtcctttgg
      901 cgtggcagca gtggcatctc tgctggccgc cctgtgctat gcagagttcg gagcaagggt
      961 gccaagaacc ggaagcgcct acctgtttac atatgtctca atgggagagc tttgggcctt
     1021 cctgatcggc tggaacgtgc tgctggagta catcatcgga ggagcagcag tggcaagggc
     1081 atggtctggc tatctggaca gcatgttttc ccactctatc agaaatttca ccgagacaca
     1141 cgtgggatcc tggcaggtgc cactgctggg acactaccct gattttctgg cagcaggaat
     1201 catcctgctg gcatctgcct tcgtgagctg cggagcacgg gtgagctcct ggctgaacca
     1261 cacattcagc gccatctccc tgctggtcat cctgttcatc gtgatcctgg gcttcatcct
     1321 ggcccagcca cacaattgga gcgccgacga gggaggattt gcacctttcg gcttttctgg
     1381 cgtgatggcc ggcaccgcca gctgtttcta cgcctttgtg ggcttcgatg tgatcgcagc
     1441 atctagcgag gaggcacaga acccaaggcg ctccgtgcca ctggcaatcg caatctctct
     1501 ggcaatcgca gcaggagcct atatcctggt gtccaccgtg ctgacactga tggtgccatg
     1561 gcactccctg gacccagatt ctgccctggc agacgccttt taccagaggg gctatagatg
     1621 ggcaggcttc atcgtggcag caggatctat ctgtgccatg aatacagtgc tgctgtccct
     1681 gctgttttct ctgcccagga tcgtgtacgc aatggcagca gacggactgt tctttcaggt
     1741 gttcgcacac gtgcacccta ggacccaggt gccagtggca ggcacactgg cctttggcct
     1801 gctgaccgcc ttcctggccc tgctgctgga tctggagagc ctggtgcagt tcctgtccct
     1861 gggcaccctg ctggcctata cctttgtggc cacatccatc atcgtgctga ggttccagaa
     1921 gtcctctccc cctagctccc ctggaccagc atccccagga cctctgacaa agcagcagtc
     1981 tagcttttct gaccacctgc agctggtggg aaccgtgcac gcaagcgtgc cagagccagg
     2041 agagctgaag cctgccctga ggccatacct gggcttcctg gatggctata gcccaggagc
     2101 agtggtgaca tgggcactgg gcgtgatgct ggcatccgcc atcacaatcg gctgcgtgct
     2161 ggtgtttggc aactctaccc tgcacctgcc tcactggggc tacatcctgc tgctgctgct
     2221 gacctctgtg atgtttctgc tgagcctgct ggtgctggga gcacaccagc agcagtatcg
     2281 ggaggacctg ttccagatcc ctatggtgcc actgatcccc gccctgagca tcgtgctgaa
     2341 tatctgtctg atgctgaagc tgagctacct gacctgggtg agattttcca tctggctgct
     2401 gatgggcctg gccgtgtact tcggctatgg catcaggcac tctaaggaga accagcgcga
     2461 gctgcctggc ctgaattcta cacactatgt ggtgttccca agaggcagcc tggaggagac
     2521 cgtgcaggcc atgcagccac ccagccaggc ccccgcccag gatcctggcc acatggagta
     2581 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2641 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2701 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2761 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2821 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2881 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2941 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     3001 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     3061 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     3121 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     3181 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3241 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3301 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3361 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3421 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3481 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3541 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3601 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3661 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3721 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4441 gccttttgct cacatgtt
//
