LOCUS       sequence-259567-        3900 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 3900)
  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 3900)
  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_MFSD5
FEATURES             Location/Qualifiers
     source          1..3900
                     /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(2025..2124)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2141..2160)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2142..2160)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2165..2181)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2165..2181)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2283..2302)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2294..3103
                     /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(2361..2380)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3250..3838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3739..3758
                     /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 accatgctgg tgaccgcata cctggccttc gtgggactgc tggcatcttg
      721 cctgggactg gagctgagca ggtgtagagc aaagccacct ggaagggcct gctccaaccc
      781 atcttttctg agattccagc tggactttta ccaggtgtat ttcctggccc tggcagcaga
      841 ttggctgcag gcaccctacc tgtataagct gtaccagcac tactattttc tggagggcca
      901 gatcgccatc ctgtacgtgt gcggactggc aagcacagtg ctgttcggac tggtggcaag
      961 ctccctggtg gactggctgg gacggaagaa tagctgcgtg ctgttcagcc tgacctacag
     1021 cctgtgctgt ctgacaaagc tgtcccagga ttatttcgtg ctgctggtgg gaagagccct
     1081 gggaggactg agcaccgccc tgctgttttc cgccttcgag gcctggtaca tccacgagca
     1141 cgtggagagg cacgactttc cagccgagtg gatcccagcc acatttgcca gggcagcatt
     1201 ctggaaccac gtgctggcag tggtggcagg agtggcagca gaggccgtgg catcttggat
     1261 cggactggga cctgtggcac cattcgtggc cgccatccct ctgctggccc tggcaggcgc
     1321 cctggccctg aggaactggg gcgagaatta tgacaggcag cgcgccttta gcaggacctg
     1381 cgcaggagga ctgaggtgtc tgctgtccga tcggagagtg ctgctgctgg gcaccatcca
     1441 ggccctgttc gagtccgtga tcttcatctt cgtgttcctg tggacaccag tgctggatcc
     1501 acacggagca cctctgggaa tcatcttttc tagcttcatg gccgcctctc tgctgggctc
     1561 ctctctgtac aggatcgcca caagcaagcg gtaccacctg cagcctatgc acctgctgtc
     1621 cctggccgtg ctgatcgtgg tgttttctct gttcatgctg acctttagca catcccctgg
     1681 ccaggagtcc ccagtggagt cttttatcgc cttcctgctg atcgagctgg cctgtggcct
     1741 gtactttccc tctatgagct tcctgaggcg caaagtgatc cctgagaccg agcaggcagg
     1801 cgtgctgaac tggttcaggg tgccactgca ctccctggca tgcctgggac tgctggtgct
     1861 gcacgactct gatcggaaga ccggcacaag aaatatgttt agcatctgta gcgccgtgat
     1921 ggtcatggca ctgctggctg tcgtagggct tttcacggta gttcggcacg acgcagagct
     1981 gagagtgcca tcccccacag aggagcccta tgcccctgag ctggacccag ctttcttgta
     2041 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2101 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2161 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2221 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2281 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2341 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2401 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2461 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2521 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2581 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2641 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2701 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2761 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2821 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2881 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2941 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3001 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3061 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3121 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3181 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3241 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3301 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3361 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3421 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3481 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3541 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3601 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3661 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3721 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3781 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3841 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
