LOCUS       sequence-263272-        3921 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 3921)
  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 3921)
  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_UNC93A
FEATURES             Location/Qualifiers
     source          1..3921
                     /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"
     regulatory      1935..1944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2046..2145)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2162..2181)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2163..2181)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2186..2202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2186..2202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2304..2323)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2315..3124
                     /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(2382..2401)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3271..3859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3760..3779
                     /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 accatggaca ggtccctgcg caacgtgctg gtggtgtctt tcggctttct
      721 gctgctgttc accgcatacg gaggactgca gagcctgcag agctccctgt attccgagga
      781 gggactggga gtgaccgccc tgtccacact gtacggcggc atgctgctgt ctagcatgtt
      841 cctgccacct ctgctgatcg agcggctggg atgcaagggc accatcatcc tgagcatgtg
      901 cggctacgtg gccttttctg tgggcaattt ctttgccagc tggtataccc tgatccctac
      961 aagcatcctg ctgggactgg gagcagcacc actgtggtcc gcccagtgca cctacctgac
     1021 catcacaggc aacacacacg ccgagaaggc cggcaagaga ggcaaggaca tggtgaatca
     1081 gtatttcggc atcttctttc tgatctttca gtcctctggc gtgtggggca acctgatcag
     1141 ctccctggtg ttcggccaga ccccatccca ggagacactg ccagaggagc agctgacctc
     1201 ctgcggagcc tctgattgtc tgatggccac cacaaccaca aattctacac agcggccaag
     1261 ccagcagctg gtgtacaccc tgctgggcat ctatacaggc agcggcgtgc tggccgtgct
     1321 gatgatcgca gccttcctgc agcccatcag ggacgtgcag agggagtccg agggagagaa
     1381 gaagtctgtg cctttctgga gcaccctgct gagcaccttc aagctgtacc gggataagag
     1441 actgtgcctg ctgatcctgc tgcccctgta ttctggcctg cagcagggct ttctgtctag
     1501 cgagtacacc cggagctatg tgacctgtac actgggcatc cagttcgtgg gctacgtgat
     1561 gatctgcttt tccgccacag acgccctgtg ctctgtgctg tacggcaagg tgagccagta
     1621 taccggcaga gccgtgctgt atgtgctggg cgccgtgaca cacgtgagct gtatgatcgc
     1681 cctgctgctg tggaggccta gagcagacca cctggccgtg ttcttcgtgt tcagcggact
     1741 gtggggagtg gcagatgccg tgtggcagac ccagaacaat gccctgtacg gcgtgctgtt
     1801 cgagaagtct aaggaggccg cctttgccaa ctataggctg tgggaggccc tgggcttcgt
     1861 gatcgccttt ggctacagca tgtttctgtg cgtgcacgtg aagctgtata tcctgctggg
     1921 cgtgctgtcc ctgaccatgg tggcctacgg cctggtggag tgcgtggagt ctaagaaccc
     1981 aatcaggcca cacgcacctg gacaagtgaa tcaggccgag gatgaggaga tccagacaaa
     2041 gatggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2101 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2161 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2221 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2281 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2341 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2401 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2461 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2521 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2581 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2641 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2701 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2761 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2821 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2881 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2941 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3001 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3061 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3121 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3181 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3241 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3301 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3361 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3421 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3481 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3541 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3601 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3661 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3721 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3781 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3841 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3901 ctggcctttt gctcacatgt t
//
