LOCUS       sequence-261716-        4023 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 4023)
  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 4023)
  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-LETM2
FEATURES             Location/Qualifiers
     source          1..4023
                     /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(2148..2247)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2264..2283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2265..2283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2288..2304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2288..2304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2406..2425)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2417..3226
                     /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(2484..2503)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3373..3961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3862..3881
                     /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 accatggcct tctattccta caactctgtg ctggcaatcg caaggaccag
      721 gttcccaagc cactttgtgc accccacatg cagctcctat tccccatctt gtgcctttct
      781 gcacctgccc gactcccacc tgaacaagac ctgcatgaag aattatgagt ctaagaagta
      841 cagcgatcct tcccagccag gcaataccgt gctgcaccca ggcacacgcc tgatccagaa
      901 gctgcacacc tccacatgtt ggctgcagga ggtgccaggc aagcctcagc tggagcaggc
      961 aaccaagcac ccccaggtga caagccctca ggccaccaag gagacaggca tggagatcaa
     1021 ggagggcaag cagtcctatc ggcagaagat catggacgag ctgaagtact attacaacgg
     1081 cttctacctg ctgtggatcg atgcaaaggt ggcagcaagg atggtgtgga ggctgctgca
     1141 cggacaggtg ctgaccagga gagagaggcg ccggctgctg agaacatgcg tggacttctt
     1201 taggctggtg cctttcatgg tgtttctgat cgtgcccttc atggagtttc tgctgcccgt
     1261 gttcctgaag ctgtttcctg agatgctgcc aagcaccttc gagagcgagt ccaagaagga
     1321 ggagaagcag aagaagaaga tggccgtgaa gctggagctg gccaagtttc tgcaggagac
     1381 catgacagag atggccagaa ggaatagagc caagatgggc gatgcctcca cacagctgtc
     1441 tagctatgtg aagcaggtgc agaccggcca caagccctct acaaaggaga tcgtgagatt
     1501 cagcaagctg tttgaggacc agctggccct ggagcacctg gataggcctc agctggtggc
     1561 cctgtgcaag ctgctggagc tgcagacctt cggcacaaac aatctgctga ggtttcagct
     1621 gctgatgaag ctgaagtcca tcaaggccga cgatgagatc atcgccaagg agggcgtgac
     1681 cgccctgtct gtgagcgagc tgcaggcagc atgtagagca aggggaatgc gctctctggg
     1741 actgaccgag gagcagctga ggcagcagct gacagagtgg caggacctgc acctgaagga
     1801 gaacgtgccc ccttccctgc tgctgctgtc tcggaccttc tacctgatcg atgtgaagcc
     1861 aaagcccatc gagatccctc tgtctggcga ggccccaaag acagacatcc tggtggagct
     1921 gcctaccttc acagagagca aggagaatat ggtggatctg gccccacagc tgaagggcac
     1981 caaggacgag gacttcatcc agccaccccc tgtgacatcc tctccaatca ccccctctac
     2041 acctatcagc ctgccaaagg gaccaatcac cagctccgag gagcccacac tgcaggccaa
     2101 gagccagatg accgcccaga actccaaggc ctctagcaag ggcgccgacc cagctttctt
     2161 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2221 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2281 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2341 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2401 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2461 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2521 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2581 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2641 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2701 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2761 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2821 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2881 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2941 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3001 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3061 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3121 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3181 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3241 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3301 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3481 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4021 gtt
//
