LOCUS       sequence-263861-        3753 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 3753)
  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 3753)
  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_SLC35B3
FEATURES             Location/Qualifiers
     source          1..3753
                     /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(1878..1977)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1994..2013)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1995..2013)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2018..2034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2018..2034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2136..2155)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2147..2956
                     /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(2214..2233)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3103..3691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3592..3611
                     /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 accatggacc tgacccagca ggccaaggat atccagaaca tcaccgtgca
      721 ggagacaaat aagaacaata gcgagtccat cgagtgctcc aagatcacca tggacctgaa
      781 gttcaacaat tctcgcaagt acatcagcat cacagtgccc tccaagaccc agacaatgtc
      841 tcctcacatc aagagcgtgg acgatgtggt ggtgctgggc atgaacctga gcaagtttaa
      901 taagctgacc cagttcttta tctgcgtggc cggcgtgttc gtgttttatc tgatctacgg
      961 ctatctgcag gagctgatct tctccgtgga gggctttaag tcatgtggat ggtaccttac
     1021 tctggtgcag ttcgcctttt attccatctt cggcctgatc gagctgcagc tgatccagga
     1081 taagcggaga aggatcccag gcaagacata catgatcatc gcctttctga ccgtgggcac
     1141 aatgggcctg agcaacacct ccctgggcta cctgaattat cctacacagg tcatcttcaa
     1201 gtgctgtaag ctgatcccag tgatgctggg cggcgtgttt atccagggca agcggtataa
     1261 cgtggccgac gtgagcgccg ccatctgcat gtccctgggc ctgatctggt tcaccctggc
     1321 cgattctacc acagccccca actttaatct gacaggcgtg gtgctgatca gcctggccct
     1381 gtgcgcagac gccgtgatcg gcaatgtgca ggagaaggcc atgaagctgc acaacgcctc
     1441 taatagcgag atggtgctgt actcctattc tatcggcttc gtgtacatcc tgctgggact
     1501 gacctgcaca agcggactgg gaccagccgt gaccttttgt gccaagaacc ccgtgagaac
     1561 atacggctat gccttcctgt tttccctgac cggctatttc ggcatctctt ttgtgctggc
     1621 cctgatcaag atcttcggcg ccctgatcgc agtgaccgtg accacaggca ggaaggccat
     1681 gacaatcgtg ctgtccttta tcttctttgc caagcccttc acatttcagt acgtgtggtc
     1741 tggcctgctg gtggtgctgg gcatcttcct gaacgtgtac agcaagaata tggacaagat
     1801 ccggctgcct tccctgtatg atctgatcaa taagtctgtg gaggcaagga agagccggac
     1861 cctggcacag acagtggacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1921 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1981 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2041 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2101 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2161 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2221 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2281 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2341 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2401 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2461 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2521 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2581 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2641 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2701 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2761 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2821 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2881 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2941 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3001 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3061 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3121 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3181 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3241 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3301 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3361 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3421 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3481 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3541 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3601 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3661 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3721 cctggccttt tgctggcctt ttgctcacat gtt
//
