LOCUS       sequence-348933-        4074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4074)
  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 4074)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..4074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1179..1190)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1631..1730)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1747..1766)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1748..1766)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1771..1787)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1771..1787)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1889..1908)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1900..2709
                     /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(1967..1986)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2856..3444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3345..3364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3598..3615
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3774..3801)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3893..3979)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4028..4050
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4042..4059
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4043..4059
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cggcgccggc
      121 cctaagagga gagccctggc agcaccagca gcagaggaga aggaggaggc aagggagaag
      181 atgctggcag caaagtccgc cgacggatct gccccagcag gagagggcga gggcgtgacc
      241 ctgcagcgca acatcacact gctgaatggc gtggccatca tcgtgggcac catcatcggc
      301 tccggcatct tcgtgacccc aacaggcgtg ctgaaggagg ccggctctcc aggactggcc
      361 ctggtggtgt gggcagcatg cggcgtgttt agcatcgtgg gcgccctgtg ctacgcagag
      421 ctgggcacca caatcagcaa gtccggcggc gattacgcct atatgctgga ggtgtatggc
      481 agcctgcccg ccttcctgaa gctgtggatc gagctgctga tcatccggcc tagctcccag
      541 tacatcgtgg ccctggtgtt cgccacctat ctgctgaagc ctctgtttcc aacatgccca
      601 gtgcctgagg aggcagcaaa gctggtggca tgcctgtgcg tgctgctgct gaccgccgtg
      661 aactgttact ccgtgaaggc cgccacaaga gtgcaggacg ccttcgccgc agcaaagctg
      721 ctggccctgg ccctgatcat cctgctgggc tttgtgcaga tcggcaaggg cgacgtgtct
      781 aacctggatc ctaatttcag ctttgagggc accaagctgg atgtgggcaa catcgtgctg
      841 gccctgtact ctggcctgtt cgcctacggc ggctggaact atctgaattt tgtgacagag
      901 gagatgatca acccctatag gaatctgcct ctggccatca tcatcagcct gcctatcgtg
      961 accctggtgt acgtgctgac aaatctggcc tatttcacca cactgtccac cgagcagatg
     1021 ctgtctagcg aggccgtggc cgtggacttt ggcaactacc acctgggcgt gatgagctgg
     1081 atcatcccag tgttcgtggg cctgagctgc tttggctccg tgaatggctc tctgttcacc
     1141 tcctctcggc tgttctttgt gggctctaga gagggccacc tgccctctat cctgagcatg
     1201 atccaccctc agctgctgac accagtgccc agcctggtgt tcacctgcgt gatgacactg
     1261 ctgtatgcct tcagcaagga tatcttttcc gtgatcaact tcttttcctt ctttaattgg
     1321 ctgtgcgtgg ccctggccat catcggcatg atctggctga ggcacaggaa gccagagctg
     1381 gagaggccca tcaaggtgaa cctggccctg ccagtgttct ttatcctggc ctgcctgttc
     1441 ctgatcgccg tgtccttttg gaagaccccc gtggagtgtg gcatcggctt cacaatcatc
     1501 ctgtctggcc tgcccgtgta cttcttcggc gtgtggtgga agaataagcc aaagtggctg
     1561 ctgcagggca tcttttccac caccgtgctg tgccagaagc tgatgcaggt ggtgccccag
     1621 gagacctaag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1681 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1741 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     1801 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     1861 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     1921 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1981 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     2041 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     2101 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     2161 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     2221 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     2281 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     2341 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     2401 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     2461 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     2521 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2581 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2641 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2701 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     2761 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     2821 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2881 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2941 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3001 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3061 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3121 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3181 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3241 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3301 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3361 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3421 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3481 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     3541 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3601 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     3661 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3721 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     3781 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     3841 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     3901 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     3961 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac gctagcatgg
     4021 atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg gccc
//
