LOCUS       sequence-472138-        3597 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 3597)
  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 3597)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..3597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(166..265)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(281..298)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(281..297)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(290..312)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      361..447
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      539..566
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(725..742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(896..1484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(976..995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1631..2440)
                     /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     2354..2373
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2432..2451
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2553..2569
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2553..2569
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2574..2593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2574..2592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2610..2709
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ccaccttctc ggccagcagg ttctccttct tcttctggcc tcctcccagc tggatgcaca
       61 gcagcagcag taataacagg aagggaaaag agccggttgg caggtaccgc agccggcggc
      121 cggcctgcct tctccgggat ggtgctcccc ttgctcccat ggtggagcct gcttttttgt
      181 acaaagttgg cattataaaa aagcattgct catcaatgtg ttgcaacgaa caggtcacta
      241 tcagtcaaaa taaaatcatt atttggggcc cgagcttaag actggccgtc gttttacaac
      301 gtcgtgactg ggaaaacatc catgctagcg ttaacgcgag agtagggaac tgccaggcat
      361 caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg
      421 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa
      481 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag
      541 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgg ctagcggtac
      601 gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
      661 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
      721 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      781 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      841 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1321 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1501 tttctacggg gtctgacgct cagtggaacg acgcgtaact cacgttaagg gattttggtc
     1561 atgagcttgc gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa
     1621 ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     1681 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     1741 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     1801 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     1861 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     1921 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     1981 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     2041 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2101 aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca gtggtgagta
     2161 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     2221 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     2281 gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg
     2341 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     2401 ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2461 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2521 atcagagatt ttgagacacg ggccagagct gccaggaaac agctatgacc atgtaatacg
     2581 actcactata ggggatatca gctggatggc aaataatgat tttattttga ctgatagtga
     2641 cctgttcgtt gcaacaaatt gataagcaat gctttcttat aatgccaact ttgtacaaga
     2701 aagctgggtc tcactcgaaa tccaggtcgg aataagggta gccgtgatac ttagacctga
     2761 agatggacag cagaaagctg aaaaagaaca ccaccaggcc caggcccacc agacagatga
     2821 tccttctctt gcccacgtcg cccttgcttg tggctgcctc attcagcagc accatgccca
     2881 tggtgatggc ggcgttcagc accaggatga tgtgggactc ggccacaaac tgggcctgag
     2941 aggagccgtg gatatagctc acctggccat tgtgggggtt cttgtgggcg taaggggggc
     3001 ctctgatgtg attccacatc tggccggatg tcatggcgaa cacgatgcac aggctcacca
     3061 tggcccagcc ggtcttgtta tagataaact ccagattgtt ccgcctcagg tacagcagtc
     3121 ctcccaccag gctcaccagc agggccaggg cgattgtgcc agagtaattt ggaggcctga
     3181 acactctgat gtgcacatcg gtcctgtcgg cgatccactt tgccagctgc tctgctgcga
     3241 atccgattct ctgcagatca aaggtgtctg ccctctttgg gcggcccttt ggtgggaagt
     3301 gcatgaaggt gggggcgcta ttcatgttca gctgctggaa cacgtcggtg ccctcatcat
     3361 agtccaccat agaaaagaac agcttgttac aaaaggcgct agaataccgc caggaattgg
     3421 ccaggatctg gtactcctcg tttgcctgcc tgcacacgct gcactgcctc tgaggctgca
     3481 gggctgtgaa catcacgatc atagagtaat tgcgaggggg ggccttgatg aacttcctaa
     3541 acttgtcgcc gttcattcta aagatagacc ttctggagct ccactccatc agctgct
//
