LOCUS       sequence-259479-        4437 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 4437)
  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 4437)
  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_SLC7A1
FEATURES             Location/Qualifiers
     source          1..4437
                     /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(2562..2661)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2678..2697)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2679..2697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2702..2718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2702..2718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2820..2839)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2831..3640
                     /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(2898..2917)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3787..4375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4276..4295
                     /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 accatgggct gcaaggtgct gctgaacatc ggccagcaga tgctgcggag
      721 aaaggtggtg gactgcagca gggaggagac caggctgtcc aggtgtctga atacatttga
      781 tctggtggcc ctgggagtgg gaagcaccct gggagccggc gtgtatgtgc tggcaggagc
      841 agtggcaagg gagaacgcag gccctgccat cgtgatctct tttctgatcg ccgccctggc
      901 aagcgtgctg gcaggactgt gctacggaga gttcggagca agggtgccaa agacaggatc
      961 cgcctacctg tattcttacg tgaccgtggg cgagctgtgg gccttcatca caggctggaa
     1021 tctgatcctg tcctatatca tcggcaccag ctccgtggca agggcatggt ctgccacatt
     1081 tgacgagctg atcggccggc ctatcggcga gttctctaga acccacatga cactgaacgc
     1141 ccctggcgtg ctggccgaga atccagatat ctttgccgtg atcatcatcc tgatcctgac
     1201 cggcctgctg acactgggcg tgaaggagtc cgccatggtg aacaagatct tcacctgcat
     1261 caatgtgctg gtgctgggct ttatcatggt gtctggcttc gtgaagggca gcgtgaagaa
     1321 ctggcagctg accgaggagg actttggcaa tacaagcggc cggctgtgcc tgaacaatga
     1381 taccaaggag ggcaagccag gagtgggagg cttcatgcct ttcggctttt ccggcgtgct
     1441 gtctggagca gcaacatgct tctacgcctt tgtgggcttc gactgtatcg ccaccacagg
     1501 cgaggaggtg aagaaccccc agaaggccat ccctgtgggc atcgtggcct ctctgctgat
     1561 ctgctttatc gcctatttcg gcgtgagcgc cgccctgacc ctgatgatgc catacttttg
     1621 tctggacaac aatagccccc tgcctgatgc cttcaagcac gtgggatggg agggagcaaa
     1681 gtatgcagtg gcagtgggaa gcctgtgcgc actgtctgcc agcctgctgg gatccatgtt
     1741 cccaatgccc agagtgatct acgccatggc cgaggacggc ctgctgttca agtttctggc
     1801 caacgtgaat gatcgcacca agacacccat catcgccacc ctggcatccg gagcagtggc
     1861 agccgtgatg gccttcctgt ttgacctgaa ggatctggtg gacctgatgt ccatcggcac
     1921 actgctggcc tattctctgg tggcagcatg cgtgctggtg ctgaggtatc agcctgagca
     1981 gccaaatctg gtgtaccaga tggcctccac ctctgatgag ctggacccag ccgatcagaa
     2041 cgagctggcc agcacaaatg actcccagct gggctttctg cccgaggccg agatgttctc
     2101 cctgaagacc atcctgtctc caaagaacat ggagccctct aagatcagcg gcctgatcgt
     2161 gaatatcagc acatccctga tcgccgtgct gatcatcacc ttttgcatcg tgacagtgct
     2221 gggaagggag gccctgacca agggcgccct gtgggccgtg ttcctgctgg caggaagcgc
     2281 cctgctgtgc gccgtggtga ccggcgtgat ctggagacag cccgagagca agacaaagct
     2341 gtcctttaag gtgcccttcc tgcctgtgct gccaatcctg tccatctttg tgaacgtgta
     2401 cctgatgatg cagctggatc agggaacctg ggtgcggttc gccgtgtgga tgctgattgg
     2461 ctttatcatc tatttcggct acggcctgtg gcactctgag gaggcaagcc tggacgcaga
     2521 tcaggcaaga acacctgacg gcaatctgga tcagtgcaag gacccagctt tcttgtacaa
     2581 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2641 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2701 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2761 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2821 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2881 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2941 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3001 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3061 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3121 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3181 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3241 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3301 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3361 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3421 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3481 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3541 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3601 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3661 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3721 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3781 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3841 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3901 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3961 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4021 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4081 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4141 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4201 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4261 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4321 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4381 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
