LOCUS       sequence-355836-        4065 bp    DNA     circular SYN 15-DEC-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 4065)
  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 4065)
  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-MFSD2B_STOP
FEATURES             Location/Qualifiers
     source          1..4065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1622..1721)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1738..1757)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1739..1757)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1762..1778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1762..1778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1880..1899)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1891..2700
                     /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(1958..1977)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2847..3435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3336..3355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3589..3606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3765..3792)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3884..3970)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4019..4041
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4033..4050
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4034..4050
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc agcaccacct
      121 gcacctgcag caaagggctc tccccagcct gagccacacg cccccgagcc tggcccaggc
      181 agcgccaaga ggggaaggga ggactctagg gcaggccggc tgagcttctg caccaaggtg
      241 tgctacggca tcggcggcgt gccaaaccag atcgccagct ccgccacagc cttctatctg
      301 cagctgtttc tgctggacat cgcacagatc ccagcagcac aggtgtccct ggtgctgttc
      361 ggaggcaagg tgtctggagc agcagcagat cctgtggcag gcttctttat caatagatcc
      421 cagaggaccg gctctggcag actgatgcca tgggtgctgg gctgcacacc ctttatcgcc
      481 ctggcctact tctttctgtg gttcctgcca ccctttacct ctctgagggg cctgtggtac
      541 accacattct attgcctgtt tcaggccctg gccaccttct ttcaggtgcc ctacaccgcc
      601 ctgacaatgc tgctgacacc ctgtcctaga gagagggatt ccgccaccgc ctatcggatg
      661 acagtggaga tggcaggcac cctgatggga gcaacagtgc acggactgat cgtgagcgga
      721 gcacacaggc ctcacaggtg cgaggccacc gccacaccag gaccagtgac cgtgtcccca
      781 aacgcagcac acctgtactg tatcgcagca gccgtggtgg tggtgacata tcccgtgtgc
      841 atcagcctgc tgtgcctggg agtgaaggag aggcctgacc caagcgcccc tgcatccggc
      901 ccaggcctgt ccttcctggc aggactgtct ctgaccacaa ggcaccctcc atacctgaag
      961 ctggtcatca gcttcctgtt tatctccgcc gccgtgcagg tggagcagag ctatctggtg
     1021 ctgttttgta cccacgcctc ccagctgcac gatcacgtgc agggcctggt gctgaccgtg
     1081 ctggtgtctg ccgtgctgag cacaccactg tgggagtggg tgctgcagcg gttcggcaag
     1141 aagacctccg ccttcggcat ctttgccatg gtgccctttg ccatcctgct ggcagcagtg
     1201 ccaacagcac ctgtggcata cgtggtggcc ttcgtgtctg gcgtgagcat cgccgtgagc
     1261 ctgctgctgc cctggtccat gctgcctgac gtggtggacg atttccagct gcagcacagg
     1321 cacggaccag gactggagac catcttttac tctagctacg tgttcttcac aaagctgtcc
     1381 ggagcatgcg cactgggaat ctccaccctg tctctggagt tttctggcta caaggcaggc
     1441 gtgtgcaagc aggcagagga ggtggtggtg accctgaagg tgctgatcgg cgccgtgcct
     1501 acatgcatga tcctggccgg cctgtgcatc ctgatggtgg gctctacccc aaagacaccc
     1561 agcagagatg cctcctctag gctgagcctg cggagaagga ccagctattc cctggcctaa
     1621 gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     1681 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     1741 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1801 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1861 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1921 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1981 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2041 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2101 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2161 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     2221 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2281 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2341 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2401 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2461 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2521 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2581 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2641 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2701 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2761 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2821 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2881 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2941 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3001 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3061 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3481 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3541 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3601 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3661 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3721 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3781 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3841 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3901 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3961 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4021 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
