LOCUS       sequence-265227-        4620 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 4620)
  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 4620)
  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_SLC27A5
FEATURES             Location/Qualifiers
     source          1..4620
                     /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(2745..2844)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2861..2880)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2862..2880)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2885..2901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2885..2901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3003..3022)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3014..3823
                     /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(3081..3100)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3970..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4459..4478
                     /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 accatgggcg tgaggcagca gctggccctg ttattactgc tgctgctgct
      721 gctgtgggga ctgggacagc ccgtgtggcc agtggcagtg gccctgaccc tgaggtggct
      781 gctgggcgac cctacatgct gcgtgctgct gggcctggcc atgctggccc gcccatggct
      841 gggcccctgg gtgcctcacg gactgagcct ggcagcagcc gccctggccc tgaccctgct
      901 gccagcccgg ctgccccctg gcctgagatg gctgcccgcc gatgtgatct tcctggccaa
      961 gatcctgcac ctgggcctga agatcagggg atgcctgtcc aggcagccac cagacacatt
     1021 cgtggatgcc tttgagagga gagcaagggc acagccagga agagccctgc tggtgtggac
     1081 cggacctgga gcaggatctg tgacattcgg cgagctggac gcaagggcat gccaggcagc
     1141 atgggccctg aaggcagagc tgggcgatcc tgcatccctg tgcgcaggag agccaaccgc
     1201 cctgctggtg ctggcatctc aggcagtgcc tgccctgtgc atgtggctgg gactggcaaa
     1261 gctgggatgt ccaacagcct ggatcaaccc tcacggaagg ggaatgccac tggcacacag
     1321 cgtgctgagc tccggagcac gggtgctggt ggtggaccct gatctgagag agtccctgga
     1381 ggagatcctg ccaaagctgc aggccgagaa tatcagatgt ttctacctga gccacacctc
     1441 ccctacacca ggcgtgggcg ccctgggagc cgccctggac gcagccccaa gccacccagt
     1501 gcctgcagat ctgagggcag gaatcacctg gcgctccccc gccctgttta tctatacatc
     1561 tggcaccaca ggcctgccaa agccagcaat cctgacccac gagcgggtgc tgcagatgtc
     1621 caagatgctg tctctgagcg gagcaaccgc agacgatgtg gtgtacacag tgctgcctct
     1681 gtatcacgtg atgggactgg tggtgggaat cctgggatgc ctggacctgg gagcaacctg
     1741 cgtgctggca ccaaagttct ccacatcttg cttttgggac gattgtaggc agcacggcgt
     1801 gaccgtgatc ctgtacgtgg gcgagctgct gcgctatctg tgcaacatcc cacagcagcc
     1861 cgaggacagg acccacacag tgcgcctggc catgggcaat ggcctgagag ccgacgtgtg
     1921 ggagaccttc cagcagcggt ttggccccat cagaatctgg gaggtgtacg gctctacaga
     1981 gggcaacatg ggcctggtga attatgtggg caggtgcggc gccctgggca agatgtcttg
     2041 tctgctgcgc atgctgagcc ccttcgagct ggtgcagttt gacatggagg ccgccgagcc
     2101 tgtgagggat aaccagggct tctgtatccc agtgggactg ggagagcctg gactgctgct
     2161 gaccaaggtg gtgtcccagc agccatttgt gggctaccgg ggccccagag agctgtctga
     2221 gcgcaagctg gtgaggaacg tgcgccagag cggcgacgtg tactataata caggcgacgt
     2281 gctggcaatg gatagggagg gcttcctgta ctttagggac cgcctgggcg ataccttcag
     2341 atggaagggc gagaacgtga gcacacacga ggtggagggc gtgctgtccc aggtggattt
     2401 tctgcagcag gtgaacgtgt acggcgtgtg cgtgccagga tgtgagggca aggtcggaat
     2461 ggcagcagtg cagctggcac ctggacagac cttcgacggc gagaagctgt accagcacgt
     2521 gcgggcatgg ctgcctgcat atgcaacacc acactttatc agaatccagg atgccatgga
     2581 ggtgacctct acattcaagc tgatgaagac ccggctggtg agagagggct ttaacgtggg
     2641 catcgtggtg gacccactgt tcgtgctgga taatagggca cagagctttc gcccactgac
     2701 cgcagagatg taccaggccg tgtgcgaggg cacatggagg ctggacccag ctttcttgta
     2761 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2821 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2881 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2941 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3001 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3061 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3121 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3181 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3241 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3301 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3361 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3421 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3481 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3541 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3601 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3661 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3721 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3781 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3841 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3901 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
