LOCUS       sequence-263075-        3570 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 3570)
  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 3570)
  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-SFXN5
FEATURES             Location/Qualifiers
     source          1..3570
                     /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(1695..1794)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1811..1830)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1812..1830)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1835..1851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1835..1851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1953..1972)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1964..2773
                     /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(2031..2050)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2920..3508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3409..3428
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggccg acaccgccac cacagcatcc gccgcagcag caagcgccgc
      721 atccgccagc tccgatgcac cacctttcca gctgggcaag ccaaggttcc agcagacatc
      781 tttttacggc aggttccgcc actttctgga catcatcgat cctcggaccc tgtttgtgac
      841 agagcggaga ctgagagagg ccgtgcagct gctggaggac tataagcacg gcaccctgcg
      901 gccaggcgtg acaaatgagc agctgtggtc tgcccagaag atcaagcagg ccatcctgca
      961 ccctgatacc aacgagaaga tcttcatgcc cttccggatg agcggctaca tccctttcgg
     1021 cacaccaatc gtggtgggcc tgctgctgcc caatcagacc ctggcctcta cagtgttttg
     1081 gcagtggctg aaccagagcc acaatgcctg cgtgaactac gccaacagga atgccaccaa
     1141 gccttctcca gccagcaagt tcatccaggg ctatctgggc gccgtgatct ccgccgtgtc
     1201 tatcgccgtg ggcctgaatg tgctggtgca gaaggccaac aagtttaccc ctgccacaag
     1261 gctgctgatc cagcggttcg tgccctttcc tgccgtggcc agcgccaaca tctgtaatgt
     1321 ggtgctgatg cgctatggcg agctggagga gggcatcgac gtgctggact ccgatggcaa
     1381 tctggtgggc tctagcaaga tcgcagcaag gcacgcactg ctggagaccg ccctgacaag
     1441 ggtggtgctg ccaatgccca tcctggtgct gccacccatc gtgatgagca tgctggagaa
     1501 gaccgccctg ctgcaggcaa ggccaagact gctgctgcct gtgcagtccc tggtgtgcct
     1561 ggcagccttc ggcctggccc tgccactggc catctctctg tttccccaga tgagcgagat
     1621 cgagacatcc cagctggagc cagagatcgc acaggccacc tcctctagga cagtggtgta
     1681 caacaagggc ctggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     1741 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     1801 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1861 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     1921 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     1981 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2041 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2101 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2161 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2221 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2281 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2341 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2401 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2461 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2521 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2581 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2641 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2701 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2761 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2821 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     2881 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2941 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3001 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3061 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3121 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3181 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3241 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3301 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3361 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3421 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3481 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3541 ggccttttgc tggccttttg ctcacatgtt
//
