LOCUS       sequence-253161-        4068 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 4068)
  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 4068)
  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_MFSD14B
FEATURES             Location/Qualifiers
     source          1..4068
                     /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(2193..2292)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2309..2328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2310..2328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2333..2349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2333..2349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2451..2470)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2462..3271
                     /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(2529..2548)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3418..4006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3907..3926
                     /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 accatgtccg tggagccacc tccagagctg gaggagaagg cagcatctga
      721 gccagaggca ggagccatgc ctgagaagag ggcaggagca caggcagcag gatctacatg
      781 gctgcagggc tttggcagac ccagcgtgta ccacgccgcc atcgtgatct tcctggagtt
      841 ctttgcctgg ggcctgctga ccacacctat gctgaccgtg ctgcacgaga cattttccca
      901 gcacaccttc ctgatgaacg gcctgatcca gggcgtgaag ggcctgctgt cttttctgag
      961 cgccccactg atcggcgccc tgagcgacgt gtggggccgg aagccttttc tgctgggcac
     1021 cgtgttcttt acatgcttcc caatccccct gatgagaatc tccccttggt ggtatttcgc
     1081 catgatctcc gtgtctggcg tgttcagcgt gacattttcc gtgatcttcg cctacgtggc
     1141 cgatgtgaca caggagcacg agcggagcac cgcatatgga tgggtgagcg ccacctttgc
     1201 cgcatccctg gtgagctccc cagcaatcgg agcatacctg agcgcctcct atggcgactc
     1261 tctggtggtg ctggtggcaa ccgtggtggc cctgctggat atctgcttca tcctggtggc
     1321 cgtgcctgag agcctgccag agaagatgcg gccagtgtct tggggagcac agatcagctg
     1381 gaagcaggca gacccctttg ccagcctgaa gaaggtcggc aaggattcca cagtgctgct
     1441 gatctgtatc accgtgttcc tgtcctacct gcctgaggcc ggccagtact ctagcttctt
     1501 tctgtatctg aggcaggtca tcggctttgg cagcgtgaag atcgccgcct tcatcgccat
     1561 ggtgggcatc ctgtccatcg tggcccagac agccttcctg tccatcctga tgcgctctct
     1621 gggcaacaag aataccgtgc tgctgggcct gggctttcag atgctgcagc tggcctggta
     1681 tggcttcggc tctcaggcct ggatgatgtg ggcagcagga acagtggcag caatgtcctc
     1741 tatcaccttt cccgccatct ctgccctggt gagcaggaac gcagagagcg accagcaggg
     1801 agtggcacag ggcatcatca caggaatcag gggactgtgc aatggactgg gacctgccct
     1861 gtacggcttc atcttttata tgttccacgt ggagctgacc gagctgggcc caaagctgaa
     1921 ctccaacaat gtgccactgc agggagccgt gatcccagga ccacctttcc tgtttggagc
     1981 atgtatcgtg ctgatgtctt ttctggtggc cctgttcatc ccagagtact ccaaggcctc
     2041 tggcgtgcag aagcactcca acagctcctc tggctctctg accaatacac ctgagagagg
     2101 cagcgacgag gatatcgagc cactgctgca ggatagctcc atctgggagc tgtctagctt
     2161 cgaggagccc ggcaatcagt gtaccgagct ggacccagct ttcttgtaca aagttggcat
     2221 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2281 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2341 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2401 aataatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2461 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2521 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2581 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2641 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2701 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2761 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2821 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2881 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2941 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3001 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3061 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3121 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3181 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3241 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3301 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3361 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
