LOCUS       sequence-261168-        4071 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 4071)
  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 4071)
  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_SLC7A5
FEATURES             Location/Qualifiers
     source          1..4071
                     /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"
     CDS             complement(1747..1758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2196..2295)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2312..2331)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2313..2331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2336..2352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2336..2352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2454..2473)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2465..3274
                     /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(2532..2551)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3421..4009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3910..3929
                     /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 accatggccg gcgccggccc taagaggaga gccctggcag caccagcagc
      721 agaggagaag gaggaggcaa gggagaagat gctggcagca aagtccgccg acggatctgc
      781 cccagcagga gagggcgagg gcgtgaccct gcagcgcaac atcacactgc tgaatggcgt
      841 ggccatcatc gtgggcacca tcatcggctc cggcatcttc gtgaccccaa caggcgtgct
      901 gaaggaggcc ggctctccag gactggccct ggtggtgtgg gcagcatgcg gcgtgtttag
      961 catcgtgggc gccctgtgct acgcagagct gggcaccaca atcagcaagt ccggcggcga
     1021 ttacgcctat atgctggagg tgtatggcag cctgcccgcc ttcctgaagc tgtggatcga
     1081 gctgctgatc atccggccta gctcccagta catcgtggcc ctggtgttcg ccacctatct
     1141 gctgaagcct ctgtttccaa catgcccagt gcctgaggag gcagcaaagc tggtggcatg
     1201 cctgtgcgtg ctgctgctga ccgccgtgaa ctgttactcc gtgaaggccg ccacaagagt
     1261 gcaggacgcc ttcgccgcag caaagctgct ggccctggcc ctgatcatcc tgctgggctt
     1321 tgtgcagatc ggcaagggcg acgtgtctaa cctggatcct aatttcagct ttgagggcac
     1381 caagctggat gtgggcaaca tcgtgctggc cctgtactct ggcctgttcg cctacggcgg
     1441 ctggaactat ctgaattttg tgacagagga gatgatcaac ccctatagga atctgcctct
     1501 ggccatcatc atcagcctgc ctatcgtgac cctggtgtac gtgctgacaa atctggccta
     1561 tttcaccaca ctgtccaccg agcagatgct gtctagcgag gccgtggccg tggactttgg
     1621 caactaccac ctgggcgtga tgagctggat catcccagtg ttcgtgggcc tgagctgctt
     1681 tggctccgtg aatggctctc tgttcacctc ctctcggctg ttctttgtgg gctctagaga
     1741 gggccacctg ccctctatcc tgagcatgat ccaccctcag ctgctgacac cagtgcccag
     1801 cctggtgttc acctgcgtga tgacactgct gtatgccttc agcaaggata tcttttccgt
     1861 gatcaacttc ttttccttct ttaattggct gtgcgtggcc ctggccatca tcggcatgat
     1921 ctggctgagg cacaggaagc cagagctgga gaggcccatc aaggtgaacc tggccctgcc
     1981 agtgttcttt atcctggcct gcctgttcct gatcgccgtg tccttttgga agacccccgt
     2041 ggagtgtggc atcggcttca caatcatcct gtctggcctg cccgtgtact tcttcggcgt
     2101 gtggtggaag aataagccaa agtggctgct gcagggcatc ttttccacca ccgtgctgtg
     2161 ccagaagctg atgcaggtgg tgccccagga gaccgaccca gctttcttgt acaaagttgg
     2221 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2281 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2341 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2401 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2461 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2521 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2581 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2641 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2701 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2761 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2821 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2881 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2941 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     3001 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3061 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3121 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3181 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3241 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3301 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3361 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3541 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3781 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
