LOCUS       sequence-479338-        3756 bp    DNA     circular SYN 28-AUG-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 3756)
  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 3756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 28, 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-SLC35B3_STOP
FEATURES             Location/Qualifiers
     source          1..3756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     121..140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     374..391
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(550..577)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(669..755)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     804..826
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     818..835
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     819..835
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    851..950
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2163..2262)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2279..2298)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2280..2298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2303..2319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2303..2319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2421..2440)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2432..3241
                     /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(2499..2518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3388..3756,1..220)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
       61 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      241 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      301 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
      361 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
      421 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      481 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
      541 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
      601 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
      661 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
      721 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
      781 gttaacgcta gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa
      841 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat
      901 tgatgagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga
      961 cctgacccag caggccaagg atatccagaa catcaccgtg caggagacaa ataagaacaa
     1021 tagcgagtcc atcgagtgct ccaagatcac catggacctg aagttcaaca attctcgcaa
     1081 gtacatcagc atcacagtgc cctccaagac ccagacaatg tctcctcaca tcaagagcgt
     1141 ggacgatgtg gtggtgctgg gcatgaacct gagcaagttt aataagctga cccagttctt
     1201 tatctgcgtg gccggcgtgt tcgtgtttta tctgatctac ggctatctgc aggagctgat
     1261 cttctccgtg gagggcttta agtcatgtgg atggtacctt actctggtgc agttcgcctt
     1321 ttattccatc ttcggcctga tcgagctgca gctgatccag gataagcgga gaaggatccc
     1381 aggcaagaca tacatgatca tcgcctttct gaccgtgggc acaatgggcc tgagcaacac
     1441 ctccctgggc tacctgaatt atcctacaca ggtcatcttc aagtgctgta agctgatccc
     1501 agtgatgctg ggcggcgtgt ttatccaggg caagcggtat aacgtggccg acgtgagcgc
     1561 cgccatctgc atgtccctgg gcctgatctg gttcaccctg gccgattcta ccacagcccc
     1621 caactttaat ctgacaggcg tggtgctgat cagcctggcc ctgtgcgcag acgccgtgat
     1681 cggcaatgtg caggagaagg ccatgaagct gcacaacgcc tctaatagcg agatggtgct
     1741 gtactcctat tctatcggct tcgtgtacat cctgctggga ctgacctgca caagcggact
     1801 gggaccagcc gtgacctttt gtgccaagaa ccccgtgaga acatacggct atgccttcct
     1861 gttttccctg accggctatt tcggcatctc ttttgtgctg gccctgatca agatcttcgg
     1921 cgccctgatc gcagtgaccg tgaccacagg caggaaggcc atgacaatcg tgctgtcctt
     1981 tatcttcttt gccaagccct tcacatttca gtacgtgtgg tctggcctgc tggtggtgct
     2041 gggcatcttc ctgaacgtgt acagcaagaa tatggacaag atccggctgc cttccctgta
     2101 tgatctgatc aataagtctg tggaggcaag gaagagccgg accctggcac agacagtgta
     2161 agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2221 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2281 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2341 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2401 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2461 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2521 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2581 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2641 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2701 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2761 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2821 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2881 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2941 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3001 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3061 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3121 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3181 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3241 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3301 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3721 cacacagccc agcttggagc gaacgaccta caccga
//
