LOCUS       sequence-262974-        3594 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 3594)
  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 3594)
  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-TUSC3
FEATURES             Location/Qualifiers
     source          1..3594
                     /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(1719..1818)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1835..1854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1836..1854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1859..1875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1977..1996)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1988..2797
                     /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(2055..2074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2944..3532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3433..3452
                     /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 accatgggag caaggggagc accatcccgg agaaggcagg ccggccgccg
      721 gctgcggtac ctgccaaccg gctcttttcc cttcctgtta ttactgctgc tgctgtgcat
      781 ccagctggga ggaggccaga agaagaagga gaacctgctg gccgagaagg tggagcagct
      841 gatggagtgg agctccagaa ggtctatctt tagaatgaac ggcgacaagt ttaggaagtt
      901 catcaaggcc ccccctcgca attactctat gatcgtgatg ttcacagccc tgcagcctca
      961 gaggcagtgc agcgtgtgca ggcaggcaaa cgaggagtac cagatcctgg ccaattcctg
     1021 gcggtattct agcgcctttt gtaacaagct gttcttttct atggtggact atgatgaggg
     1081 caccgacgtg ttccagcagc tgaacatgaa tagcgccccc accttcatgc acttcccacc
     1141 aaagggccgc ccaaagaggg cagacacctt tgatctgcag agaatcggat tcgcagcaga
     1201 gcagctggca aagtggatcg ccgacaggac cgatgtgcac atcagagtgt tcaggcctcc
     1261 aaattactct ggcacaatcg ccctggccct gctggtgagc ctggtgggag gactgctgta
     1321 cctgaggcgg aacaatctgg agtttatcta taacaagacc ggctgggcca tggtgagcct
     1381 gtgcatcgtg ttcgccatga catccggcca gatgtggaat cacatcagag gcccccctta
     1441 cgcccacaag aacccccaca atggccaggt gagctatatc cacggctcct ctcaggccca
     1501 gtttgtggcc gagtcccaca tcatcctggt gctgaacgcc gccatcacca tgggcatggt
     1561 gctgctgaat gaggcagcca caagcaaggg cgacgtgggc aagagaagga tcatctgtct
     1621 ggtgggcctg ggcctggtgg tgttcttttt cagctttctg ctgtccatct tcaggtctaa
     1681 gtatcacggc tacccttatt ccgacctgga tttcgaggac ccagctttct tgtacaaagt
     1741 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1801 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1861 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1921 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1981 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2041 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2101 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2161 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2221 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2281 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2341 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2401 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2461 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2521 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2581 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2641 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2701 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2761 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2821 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2881 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
