LOCUS       sequence-265776-        4488 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 4488)
  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 4488)
  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_SLC27A1
FEATURES             Location/Qualifiers
     source          1..4488
                     /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(2613..2712)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2729..2748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2730..2748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2753..2769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2753..2769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2871..2890)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2882..3691
                     /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(2949..2968)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3838..4426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4327..4346
                     /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 accatgaggg cacctggagc aggagcagca tccgtggtgt ctctggccct
      721 gctgtggctg ctgggactgc catggacctg gtccgccgca gccgccctgg gcgtgtatgt
      781 gggatctgga ggatggaggt tcctgaggat cgtgtgcaag acagcccgga gagatctgtt
      841 tggcctgagc gtgctgatca gggtgcgcct ggagctgagg cgccaccaga gggccggcca
      901 caccatcccc agaatcttcc aggcagtggt gcagaggcag cctgagagac tggccctggt
      961 ggatgcagga accggagagt gttggacatt tgcccagctg gacgcctact ccaacgccgt
     1021 ggccaatctg ttccggcagc tgggatttgc accaggcgac gtggtggcca tcttcctgga
     1081 gggcagaccc gagtttgtgg gactgtggct gggactggca aaggcaggaa tggaggccgc
     1141 cctgctgaac gtgaatctgc ggagagagcc tctggcattc tgcctgggca cctccggagc
     1201 aaaggccctg atctttggag gagagatggt ggcagcagtg gcagaggtga gcggacacct
     1261 gggcaagtcc ctgatcaagt tctgttctgg cgacctggga cctgagggaa tcctgccaga
     1321 cacccacctg ctggatcctc tgctgaagga ggcaagcaca gcaccactgg cacagatccc
     1381 ttccaaggga atggacgata ggctgttcta catctatacc agcggaacca caggactgcc
     1441 aaaggcagcc atcgtggtgc actcccggta ctatagaatg gccgcctttg gccaccacgc
     1501 atataggatg caggcagcag acgtgctgta cgattgcctg cccctgtatc actctgccgg
     1561 caacatcatc ggcgtgggcc agtgtctgat ctacggcctg accgtggtgc tgaggaagaa
     1621 gttcagcgcc tcccgctttt gggacgattg catcaagtac aattgtacag tggtgcagta
     1681 tatcggcgag atctgccggt acctgctgaa gcagccagtg agagaggcag agaggcgcca
     1741 ccgggtgaga ctggcagtgg gaaacggact gaggccagca atctgggagg agttcaccga
     1801 gaggtttggc gtgcgccaga tcggcgagtt ctatggcgcc acagagtgca actgtagcat
     1861 cgccaatatg gatggcaagg tcggctcctg tggctttaac tctaggatcc tgccccacgt
     1921 gtaccctatc cgcctggtga aagtgaatga ggataccatg gagctgctga gagacgccca
     1981 gggcctgtgc atcccatgtc aggcaggaga gccaggactg ctggtgggac agatcaatca
     2041 gcaggacccc ctgcggagat tcgatggcta cgtgagcgag agcgccacat ccaagaagat
     2101 cgcccacagc gtgttttcca agggcgattc tgcctacctg agcggcgacg tgctggtcat
     2161 ggatgagctg ggctacatgt atttcaggga ccgctctggc gatacctttc ggtggagagg
     2221 cgagaacgtg tctaccacag aggtggaggg cgtgctgagc aggctgctgg gacagacaga
     2281 cgtggccgtg tacggcgtgg cagtgcctgg agtggagggc aaggcaggca tggccgccgt
     2341 ggccgaccct cacagcctgc tggatccaaa tgccatctat caggagctgc agaaggtgct
     2401 ggcaccatac gcaaggccca tcttcctgag actgctgcca caggtggaca ccacaggcac
     2461 cttcaagatc cagaagacaa ggctgcagag ggagggattt gacccaaggc agacctctga
     2521 tcgcctgttc tttctggacc tgaagcaggg ccactatctg cctctgaacg aggccgtgta
     2581 cacacgcatc tgcagcggcg cctttgccct ggacccagct ttcttgtaca aagttggcat
     2641 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2701 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2761 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2821 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2881 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2941 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3001 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3061 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3121 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3181 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3241 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3301 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3361 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3421 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3481 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3541 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3601 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3661 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3721 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3781 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3961 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4021 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4081 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4141 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4201 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4261 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4321 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4381 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4441 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
