LOCUS       sequence-408928-        3849 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 3849)
  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 3849)
  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-SLC35B2_STOP
FEATURES             Location/Qualifiers
     source          1..3849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1406..1505)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1522..1541)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1523..1541)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1546..1562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1546..1562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1664..1683)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1675..2484
                     /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(1742..1761)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2631..3219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3120..3139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3373..3390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3549..3576)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3668..3754)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3803..3825
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3817..3834
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3818..3834
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga cgcaaggtgg
      121 tgggcagtgg tggtgctggc cgccttccct agcctgggag caggaggaga gaccccagag
      181 gcaccacctg agtcctggac acagctgtgg ttctttaggt tcgtggtgaa cgcagcagga
      241 tacgcaagct ttatggtgcc cggctacctg ctggtgcagt attttcggag aaagaactat
      301 ctggagaccg gaaggggact gtgcttccca ctggtgaagg cctgcgtgtt cggcaatgag
      361 cctaaggcat ccgacgaggt gcctctggca ccaagaacag aggccgccga gaccacacct
      421 atgtggcagg ccctgaagct gctgttctgc gccaccggcc tgcaggtgtc ctacctgaca
      481 tggggcgtgc tgcaggagag agtgatgacc cgcagctatg gagcaaccgc cacatcccca
      541 ggagagaggt tcacagattc tcagtttctg gtgctgatga accgcgtgct ggccctgatc
      601 gtggcaggac tgtcttgcgt gctgtgcaag cagccaaggc acggagcacc aatgtaccgc
      661 tatagcttcg cctctctgag caatgtgctg agctcctggt gtcagtacga ggccctgaag
      721 ttcgtgtcct ttccaaccca ggtgctggcc aaggcctcta aagtgatccc cgtgatgctg
      781 atgggcaagc tggtgtccag gcgctcttac gagcactggg agtatctgac cgccacactg
      841 atctccatcg gcgtgtctat gttcctgctg tctagcggac cagagccacg gtcctctcct
      901 gcaaccacac tgagcggcct gatcctgctg gccggctaca tcgccttcga cagctttacc
      961 tccaactggc aggatgccct gtttgcctat aagatgagct ccgtgcagat gatgttcggc
     1021 gtgaatttct tttcttgcct gtttaccgtg ggaagcctgc tggagcaggg cgccctgctg
     1081 gagggcacac ggttcatggg cagacactcc gagtttgccg cccacgccct gctgctgtct
     1141 atctgcagcg cctgtggcca gctgttcatc ttttacacca tcggccagtt cggcgccgcc
     1201 gtgtttacca tcatcatgac actgcggcag gccttcgcca tcctgctgtc ttgtctgctg
     1261 tacggccaca ccgtgacagt ggtgggagga ctgggagtgg cagtggtgtt tgccgccctg
     1321 ctgctgcggg tgtatgcaag gggcagactg aagcagagag gcaagaaggc cgtgcccgtg
     1381 gagtcccctg tgcagaaggt gtaagaccca gctttcttgt acaaagttgg cattataaga
     1441 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1501 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1561 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1621 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1681 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1741 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     1801 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1861 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1921 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     1981 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2041 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2101 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2161 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2221 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2281 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2341 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2401 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2461 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2521 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2581 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3241 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3301 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3361 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3421 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3481 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     3541 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3601 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3661 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3721 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     3781 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     3841 ctcgggccc
//
