LOCUS       sequence-259490-        4104 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 4104)
  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 4104)
  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_MFSD4B
FEATURES             Location/Qualifiers
     source          1..4104
                     /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(2229..2328)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2345..2364)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2346..2364)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2369..2385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2369..2385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2487..2506)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2498..3307
                     /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(2565..2584)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3454..4042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3943..3962
                     /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 accatgctgt gcgcaagctt cctgggactg ggactgtccg tggcaatcgt
      721 gggccccacc tttcaggatc tggccacaaa cgtgaatagg aacatcagct ccctgagctt
      781 catctttgtg ggacgcgccc tgggatacct gagcggatcc gtgatcggag gcttcctggt
      841 ggacgtgatg aattattttc tgctgctggg catctctatg agcgccacca cagtgggcct
      901 gtacctggtg ccattctgta agaccgccat cctgctgaca gtgatgatga gcatctttgg
      961 cgtgtccatc ggcatcctgg ataccggcgg caacgtgctg atcctggcaa tctggggcga
     1021 caagggagca ccacacatgc aggccctgca cttctccttt gccctgggag ccttcctggc
     1081 accactgctg gcaaagctgg ccctgggacc aaccgcctct gccgagaatc acacagagag
     1141 cgatttccac cctgccctga accagtctag cgacgccgat agcgaggccc tgtttggcgt
     1201 gccaaatgac aagaacctgc tgtgggccta cgccgtgatc ggcacctata tgttcctggt
     1261 gtccgtgatc ttcttttgcc tgtttctgaa gaattcctct aagcaggaga aggcaagggc
     1321 aagcgccgag accttccgga gagccaagta ccacaacgcc ctgctgtgcc tgctgtttct
     1381 gttctttttc ttttatgtgg gcgccgaggt gacatacggc agctacgtgt tcagcttcgc
     1441 caccacacac gcaggaatga aggagtccga ggcagcaggc ctgaattcta tcttctgggg
     1501 cacctttgcc gcatgcagag gactggccat cttctttgcc acctgtctgc agcctggcac
     1561 aatgatcgtg ctgtctaata tcggcagcct gaccagctcc ctgttcctgg tgctgtttga
     1621 taagaacccc atctgcctgt ggatcgccac cagcgtgtac ggagcaagca tggccaccac
     1681 attcccttcc ggcgtgtctt ggatcgagca gtacaccaca atccacggca agagcgccgc
     1741 cttctttgtg atcggagcat ccctgggaga gatggcaatc ccagccgtga tcggcatcct
     1801 gcagggcaag taccccgatc tgcctgtggt gctgtatacc tccctgggcg cctctatcgc
     1861 cacaggcatc ctgttccctg tgctgtataa gctggccacc tctccactgg acaggcagcg
     1921 caaggaggac aggaagagcg aggatcagaa ggccctgctg tctagctccg gcctgaatga
     1981 gtacgaggag gagaacgagg aggaggacgc cgagaagtgg aatgagatgg attttgagat
     2041 gatcgagacc aacgacacaa tgcggcactc catcatcgag acatctagat ctagcctgac
     2101 cgagcccaca gccgaggtgt acaatcagta tccttccaac gccctggtgt tcgagtcctc
     2161 tcccttcaac accggatctg cccacgtgaa gcacctgcca gagaccagga caaagggcac
     2221 aaacgtggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2281 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2341 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2401 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2461 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2521 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2581 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2641 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2701 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2761 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2821 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2881 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2941 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3001 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3061 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3121 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3181 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3241 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3301 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3361 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3601 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3841 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4081 ttgctggcct tttgctcaca tgtt
//
