LOCUS       sequence-263889-        4077 bp    DNA     circular SYN 14-APR-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 4077)
  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 4077)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SLC2A4
FEATURES             Location/Qualifiers
     source          1..4077
                     /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(2202..2301)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2318..2337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2319..2337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2342..2358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2342..2358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2460..2479)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2471..3280
                     /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(2538..2557)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3427..4015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3916..3935
                     /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 accatgcctt ctggcttcca gcagatcggc agcgaggacg gagagccacc
      721 tcagcagagg gtgaccggca cactggtgct ggccgtgttc tctgccgtgc tgggcagcct
      781 gcagtttggc tacaacatcg gcgtgatcaa tgccccacag aaagtgatcg agcagagcta
      841 taacgagacc tggctgggcc ggcagggacc agagggacct agctccatcc cacccggcac
      901 actgaccaca ctgtgggccc tgtccgtggc catcttctct gtgggcggca tgatctctag
      961 ctttctgatc ggcatcatct cccagtggct gggaaggaag agggcaatgc tggtgaacaa
     1021 tgtgctggcc gtgctgggag gatctctgat gggactggca aacgcagcag caagctacga
     1081 gatgctgatc ctgggcaggt tcctgatcgg agcatactct ggactgacca gcggactggt
     1141 gccaatgtat gtgggcgaga tcgcaccaac ccacctgagg ggcgccctgg gcacactgaa
     1201 tcagctggcc atcgtgatcg gcatcctgat cgcacaggtg ctgggactgg agagcctgct
     1261 gggaaccgca tccctgtggc ctctgctgct gggcctgaca gtgctgccag ccctgctgca
     1321 gctggtgctg ctgccatttt gccccgagag ccctaggtac ctgtatatca tccagaatct
     1381 ggagggacca gcaaggaagt ccctgaagcg cctgaccgga tgggcagacg tgtctggcgt
     1441 gctggccgag ctgaaggatg agaagcgcaa gctggagcgg gagagacccc tgtccctgct
     1501 gcagctgctg ggctctcgga cacacagaca gcctctgatc atcgccgtgg tgctgcagct
     1561 gtcccagcag ctgtctggca tcaacgccgt gttctactat tccacctcta tctttgagac
     1621 agcaggagtg ggacagccag catacgcaac catcggcgcc ggcgtggtga ataccgtgtt
     1681 cacactggtg tctgtgctgc tggtggagag ggcaggccgg agaaccctgc acctgctggg
     1741 actggcagga atgtgcggct gtgccatcct gatgacagtg gccctgctgc tgctggagag
     1801 agtgcctgcc atgagctacg tgagcatcgt ggccatcttc ggctttgtgg ccttctttga
     1861 gatcggccct ggcccaatcc cctggttcat cgtggccgag ctgttttccc agggacctag
     1921 accagcagca atggcagtgg caggctttag caactggacc tccaatttca tcatcggcat
     1981 gggctttcag tacgtggccg aggccatggg cccatacgtg ttcctgctgt ttgccgtgct
     2041 gctgctgggc ttctttatct tcacatttct gagggtgcca gagaccaggg gacgcacatt
     2101 cgatcagatc agcgccgcct ttcacagaac cccctccctg ctggagcagg aggtgaagcc
     2161 ttccacagag ctggagtatc tgggcccaga cgagaacgat gacccagctt tcttgtacaa
     2221 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2281 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2341 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2401 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2461 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2521 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2581 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2641 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2701 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2761 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2821 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     2881 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     2941 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3001 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3061 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3121 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3181 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3241 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3301 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3361 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3541 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3601 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3661 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3721 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3781 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3841 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3901 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3961 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4021 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
