LOCUS       sequence-479334-        3546 bp    DNA     circular SYN 22-JUL-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 3546)
  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 3546)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 22, 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-SLC35B4_STOP
FEATURES             Location/Qualifiers
     source          1..3546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..35)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(189..777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(269..288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(924..1733)
                     /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     1647..1666
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1725..1744
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1846..1862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1846..1862
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1903..2002
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3005..3104)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3120..3137)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3120..3136)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3129..3151)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3200..3286
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3378..3405
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
       61 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      121 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      181 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      241 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      301 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      361 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      421 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      481 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      541 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      601 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      661 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      721 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      781 gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgta actcacgtta
      841 agggattttg gtcatgagct tgcgccgtcc cgtcaagtca gcgtaatgct ctgccagtgt
      901 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
      961 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     1021 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     1081 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     1141 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     1201 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     1261 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     1321 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     1381 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc
     1441 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     1501 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     1561 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag
     1621 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     1681 tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct cataacaccc
     1741 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     1801 tgtgcaatgt aacatcagag attttgagac acgggccaga gctgccagga aacagctatg
     1861 accatgtaat acgactcact ataggggata tcagctggat ggcaaataat gattttattt
     1921 tgactgatag tgacctgttc gttgcaacaa attgataagc aatgctttct tataatgcca
     1981 actttgtaca agaaagctgg gtcttaattc ttcttagagt ccttctgggg ctcggacttt
     2041 gtggtgccca gattgttcca cacttctgta tacatcaggg tgccgataaa cacgaacagt
     2101 gtgcccagcc agtgccacag ggtaaaaggg ttctggaagt acaggataga aaagatcagg
     2161 gacacgaact tgcgcagggt caccaccagt gtcacggtca ggctggcaca ctctgtggtc
     2221 aggataaaca cgcccctgat gcacacgtac tgtgtgatga tgttcatcag caggtagaac
     2281 cacatgattg gcagggtcac gccgatcaca gggatctcat acagctcgga cttattaaac
     2341 agcacggcgt gatcgtagat gtcgctggcc agaaacacga agccgggcag tggcagggcg
     2401 tggttataga acagggcctc cttgctgtgc ttgccgaacc gcttgtacag tgtctcctgg
     2461 aagatgccca tcctggcaga catcagcagg gcgaaggtca gggcgccgat tcccagcagc
     2521 caccacacga aggcctggaa gccatcattc tcgctcaggg agctctgaga tgtcacctgc
     2581 ttggcagaca taaaggtaca gatgaagatg cccacggaca ccagggcgat gcttgtatac
     2641 ttaaagatgg agtatctctt cttcaggatg atgatgccca ggatcatgtt ggcgatcagg
     2701 ctgccagacc ggaagatcat gtgcaggggc atggcgatat tcaggttcag ggcgtaattg
     2761 ttcaccacgg acactgtaaa gaacatggtc accatgatgg catagtatct gattgggatt
     2821 gcaggtggct tccttcccag gtctgcctca aacaggaagc cctccacggc gataaacagg
     2881 aactgggcga aggtcacgat attgccgcat ccagggtgct tccttgccag cagctccaga
     2941 aagatcacgt tgctacagca gccggcgaac accagtccca ctgccagggc gggcctcatg
     3001 gtggagcctg cttttttgta caaagttggc attataaaaa agcattgctc atcaatgtgt
     3061 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc gagcttaaga
     3121 ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt taacgcgaga
     3181 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg
     3241 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     3301 tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     3361 caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     3421 tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cactgcccgc tttccagtcg
     3481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3541 cgtatt
//
