LOCUS       sequence-263139-        4341 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 4341)
  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 4341)
  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_UNC93B1
FEATURES             Location/Qualifiers
     source          1..4341
                     /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(2466..2565)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2582..2601)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2583..2601)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2606..2622)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2606..2622)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2724..2743)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2735..3544
                     /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(2802..2821)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3691..4279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4180..4199
                     /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 accatggagg ccgagccccc tctgtaccca atggcaggag cagcaggacc
      721 tcagggcgac gaggatctgc tgggagtgcc tgatggacca gaggcaccac tggacgagct
      781 ggtgggagca taccccaact ataatgagga ggaggaggag cggagatact ataggcgcaa
      841 gagactgggc gtgctgaaga acgtgctggc agcatctgcc ggaggaatgc tgacctacgg
      901 cgtgtatctg ggcctgctgc agatgcagct gatcctgcac tatgatgaga catacaggga
      961 ggtgaagtat ggcaacatgg gcctgcctga catcgatagc aagatgctga tgggcatcaa
     1021 tgtgacccca atcgccgccc tgctgtacac acccgtgctg atcagattct ttggcaccaa
     1081 gtggatgatg ttcctggccg tgggcatcta tgccctgttt gtgagcacca actactggga
     1141 gaggtactat acactggtgc cttccgccgt ggccctggga atggcaatcg tgccactgtg
     1201 ggcctccatg ggcaattaca tcacacgcat ggcccagaag taccacgagt attctcacta
     1261 caaggagcag gatggacagg gaatgaagca gaggccacca aggggatccc acgcaccata
     1321 tctgctggtg ttccaggcca tcttttacag cttctttcac ctgtccttcg cctgcgcaca
     1381 gctgccaatg atctattttc tgaaccacta cctgtatgac ctgaaccaca ccctgtacaa
     1441 tgtgcagtct tgtggcacaa acagccacgg catcctgtcc ggcttcaata agaccgtgct
     1501 gagaacactg cctaggtctg gcaatctgat cgtggtggag agcgtgctga tggccgtggc
     1561 ctttctggcc atgctgctgg tgctgggact gtgcggagca gcctatcgcc caaccgagga
     1621 gatcgatctg cggagcgtgg gctggggcaa tatcttccag ctgcccttta agcacgtgcg
     1681 cgactatcgg ctgagacacc tggtgccttt ctttatctac tccggcttcg aggtgctgtt
     1741 tgcatgcaca ggaatcgccc tgggatacgg cgtgtgctct gtgggactgg agcggctggc
     1801 ctatctgctg gtggcctaca gcctgggcgc ctctgccgcc agcctgctgg gactgctggg
     1861 actgtggctg ccaaggcctg tgccactggt ggccggcgcc ggcgtgcacc tgctgctgac
     1921 cttcatcctg ttcttttggg caccagtgcc tcgggtgctg cagcactcct ggatcctgta
     1981 tgtggcagcc gccctgtggg gagtgggatc cgccctgaac aagaccggcc tgtctacact
     2041 gctgggcatc ctgtacgagg ataaggagag acaggacttc atctttacca tctatcactg
     2101 gtggcaggcc gtggccatct tcacagtgta cctgggcagc tccctgcaca tgaaggccaa
     2161 gctggccgtg ctgctggtga ccctggtggc agccgccgtg agctacctga ggatggagca
     2221 gaagctgagg agaggagtgg cacctaggca gccaagaatc ccaaggccac agcacaaggt
     2281 gcgcggatac aggtatctgg aggaggacaa ttccgatgag tctgacgccg agggcgagca
     2341 cggcgatggc gccgaggagg aggcccctcc agccggccct aggccaggac cagagcctgc
     2401 aggactgggc aggcgcccat gtccatacga gcaggcacag ggcggcgacg gccccgagga
     2461 gcaggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2521 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2581 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2641 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2701 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2761 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2821 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2881 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2941 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3001 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3061 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3121 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3181 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3241 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3301 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3361 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3421 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3481 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3541 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3601 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3661 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3721 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3781 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3841 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3901 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3961 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4021 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4081 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4141 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4201 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4261 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4321 ctggcctttt gctcacatgt t
//
