LOCUS       sequence-263655-        4062 bp    DNA     circular SYN 15-APR-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 4062)
  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 4062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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
FEATURES             Location/Qualifiers
     source          1..4062
                     /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(2187..2286)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2303..2322)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2304..2322)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2327..2343)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2327..2343)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2445..2464)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2456..3265
                     /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(2523..2542)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3412..4000
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3901..3920
                     /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 caccacctgc acctgcagca aagggctctc cccagcctga
      721 gccacacgcc cccgagcctg gcccaggcag cgccaagagg ggaagggagg actctagggc
      781 aggccggctg agcttctgca ccaaggtgtg ctacggcatc ggcggcgtgc caaaccagat
      841 cgccagctcc gccacagcct tctatctgca gctgtttctg ctggacatcg cacagatccc
      901 agcagcacag gtgtccctgg tgctgttcgg aggcaaggtg tctggagcag cagcagatcc
      961 tgtggcaggc ttctttatca atagatccca gaggaccggc tctggcagac tgatgccatg
     1021 ggtgctgggc tgcacaccct ttatcgccct ggcctacttc tttctgtggt tcctgccacc
     1081 ctttacctct ctgaggggcc tgtggtacac cacattctat tgcctgtttc aggccctggc
     1141 caccttcttt caggtgccct acaccgccct gacaatgctg ctgacaccct gtcctagaga
     1201 gagggattcc gccaccgcct atcggatgac agtggagatg gcaggcaccc tgatgggagc
     1261 aacagtgcac ggactgatcg tgagcggagc acacaggcct cacaggtgcg aggccaccgc
     1321 cacaccagga ccagtgaccg tgtccccaaa cgcagcacac ctgtactgta tcgcagcagc
     1381 cgtggtggtg gtgacatatc ccgtgtgcat cagcctgctg tgcctgggag tgaaggagag
     1441 gcctgaccca agcgcccctg catccggccc aggcctgtcc ttcctggcag gactgtctct
     1501 gaccacaagg caccctccat acctgaagct ggtcatcagc ttcctgttta tctccgccgc
     1561 cgtgcaggtg gagcagagct atctggtgct gttttgtacc cacgcctccc agctgcacga
     1621 tcacgtgcag ggcctggtgc tgaccgtgct ggtgtctgcc gtgctgagca caccactgtg
     1681 ggagtgggtg ctgcagcggt tcggcaagaa gacctccgcc ttcggcatct ttgccatggt
     1741 gccctttgcc atcctgctgg cagcagtgcc aacagcacct gtggcatacg tggtggcctt
     1801 cgtgtctggc gtgagcatcg ccgtgagcct gctgctgccc tggtccatgc tgcctgacgt
     1861 ggtggacgat ttccagctgc agcacaggca cggaccagga ctggagacca tcttttactc
     1921 tagctacgtg ttcttcacaa agctgtccgg agcatgcgca ctgggaatct ccaccctgtc
     1981 tctggagttt tctggctaca aggcaggcgt gtgcaagcag gcagaggagg tggtggtgac
     2041 cctgaaggtg ctgatcggcg ccgtgcctac atgcatgatc ctggccggcc tgtgcatcct
     2101 gatggtgggc tctaccccaa agacacccag cagagatgcc tcctctaggc tgagcctgcg
     2161 gagaaggacc agctattccc tggccgaccc agctttcttg tacaaagttg gcattataag
     2221 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2281 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2341 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2401 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2461 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2521 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2581 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2641 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2701 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2761 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2821 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2881 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2941 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3001 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3061 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3121 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3181 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3241 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3301 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3361 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3421 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3481 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3541 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3601 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3661 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3721 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3781 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3841 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3901 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3961 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4021 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
