LOCUS       sequence-265279-        3813 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 3813)
  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 3813)
  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_SLC35F3
FEATURES             Location/Qualifiers
     source          1..3813
                     /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(1938..2037)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2054..2073)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2055..2073)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2078..2094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2078..2094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2196..2215)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2207..3016
                     /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(2274..2293)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3163..3751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3652..3671
                     /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 accatgaaga agcactccgc cagggtggca ccactgtctg cctgcaatag
      721 ccccgtgctg accctgacaa aggtggaggg agaggagagg ccaagagact ctccaggacc
      781 tgcagaggca caggcacctg caggcgtgga ggccggcggc agggcctctc ggagatgctg
      841 gacctgtagc cgcgcccagc tgaagaagat cttctgggga gtggcagtgg tgctgtgcgt
      901 gtgcagctcc tgggcaggat ccacccagct ggccaagctg acattccgga agtttgacgc
      961 ccctttcacc ctgacatggt ttgccaccaa ctggaatttt ctgttctttc cactgtacta
     1021 tgtgggccac gtgtgcaagt ccacagagaa gcagtctgtg aagcagaggt accgcgagtg
     1081 ctgtagattc tttggcgata acggcctgac cctgaaggtg ttctttacaa aggcagcacc
     1141 tttcggcgtg ctgtggaccc tgacaaacta cctgtatctg cacgccatca agaagatcaa
     1201 taccacagac gtgtctgtgc tgttctgctg taacaaggcc ttcgtgtttc tgctgagctg
     1261 gatcgtgctg cgggatcggt tcatgggcgt gaggatcgtg gcagcaatcc tggcaatcgc
     1321 aggcatcgtg atgatgacat acgccgacgg cttccacagc cactccgtga tcggaatcgc
     1381 cctggtggtg gcatctgcca gcatgtccgc cctgtataag gtgctgttta agctgctgct
     1441 gggaagcgcc aagttcggag aggccgccct gtttctgtcc atcctgggcg tgttcaatat
     1501 cctgtttatc acctgcatcc ccatcatcct gtacttcaca aaggtggagt attggtctag
     1561 ctttgacgat atcccttggg gcaacctgtg cggcttctcc gtgctgctgc tgaccttcaa
     1621 catcgtgctg aattttggca tcgccgtgac ctacccaaca ctgatgagcc tgggcatcgt
     1681 gctgtccatc cccgtgaatg ccgtgatcga tcactataca agccagatcg tgttcaacgg
     1741 cgtgcgcgtg atcgccatca tcatcatcgg cctgggcttt ctgctgctgc tgctgccaga
     1801 ggagtgggac gtgtggctga tcaagctgct gaccaggctg aaggtgcgca agaaggagga
     1861 gccagcagag ggagcagcag atctgtcctc tggccctcag tccaagaata ggcgcgcccg
     1921 gccatctttt gccagagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1981 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2041 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2101 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2161 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2221 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2281 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2341 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2401 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2461 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2521 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2581 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2641 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2701 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2761 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2821 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2881 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2941 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3001 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3061 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3781 cctggccttt tgctggcctt ttgctcacat gtt
//
