LOCUS       sequence-265192-        3780 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 3780)
  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 3780)
  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_SLC35E1
FEATURES             Location/Qualifiers
     source          1..3780
                     /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(1905..2004)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2021..2040)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2022..2040)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2045..2061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2045..2061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2163..2182)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2174..2983
                     /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(2241..2260)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3130..3718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3619..3638
                     /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 accatggcag cagcagcagt gggagcagga cacggcgccg gcggccctgg
      721 cgccgccagc tcctctggcg gcgccaggga gggcgcccgc gtggccgccc tgtgcctgct
      781 gtggtacgca ctgagcgccg gaggcaacgt ggtgaacaaa gtgatcctga gcgccttccc
      841 atttcccgtg accgtgtccc tgtgccacat cctggccctg tgcgccggcc tgccccctct
      901 gctgagggca tggagggtgc caccagcacc tccagtgtct ggacctggac caagcccaca
      961 ccctagctcc ggaccactgc tgccacctcg gttctaccct agatatgtgc tgccactggc
     1021 cttcggcaag tactttgcca gcgtgtccgc ccacgtgtct atctggaagg tgcccgtgag
     1081 ctatgcccac accgtgaagg ccacaatgcc tatctgggtg gtgctgctga gccggatcat
     1141 catgaaggag aagcagtcca ccaaggtgta cctgtctctg atccccatca tcagcggcgt
     1201 gctgctggca accgtgacag agctgtcctt tgacatgtgg ggactggtgt ctgccctggc
     1261 agccacactg tgcttctccc tgcagaacat cttttctaag aaggtgctgc gggattccag
     1321 aatccaccac ctgaggctgc tgaatatcct gggctgtcac gccgtgttct ttatgatccc
     1381 aacctgggtg ctggtggacc tgtccgcctt cctggtgtct agcgatctga cctacgtgta
     1441 tcagtggccc tggacactgc tgctgctggc cgtgtctggc ttctgtaact ttgcccagaa
     1501 tgtgatcgcc tttagcatcc tgaacctggt gtcccctctg tcttatagcg tggccaatgc
     1561 caccaagcgg atcatggtca tcacagtgtc tctgatcatg ctgagaaacc cagtgaccag
     1621 cacaaatgtg ctgggcatga tgaccgccat cctgggcgtg ttcctgtaca acaagacaaa
     1681 gtatgacgcc aatcagcagg ccaggaagca cctgctgccc gtgaccacag ccgatctgtc
     1741 ctctaaggag cggcacagat ccccactgga gaagccccac aacggcctgc tgttccccca
     1801 gcacggcgac taccagtatg gccgcaacaa tatcctgaca gatcactttc agtacagcag
     1861 gcagtcctac cctaactcct attctctgaa tcgctatgac gtggacccag ctttcttgta
     1921 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     1981 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2041 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2101 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2161 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2221 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2281 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2341 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2401 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2461 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2521 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2581 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2641 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2701 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2761 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2821 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2881 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2941 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3001 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3061 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3241 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
