LOCUS       sequence-489473-        4023 bp    DNA     circular SYN 16-OCT-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 4023)
  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 4023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 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-MFSD14A_STOP
FEATURES             Location/Qualifiers
     source          1..4023
                     /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(2148..2247)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2264..2283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2265..2283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2288..2304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2288..2304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2406..2425)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2417..3226
                     /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(2484..2503)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3373..3961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3862..3881
                     /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 accatgaccc agggcaagaa gaagaagcgg gccgccaaca gatccatcat
      721 gctggccaag aagatcatca tcaaggacgg aggaacacca cagggaatcg gatcccctag
      781 cgtgtaccac gccgtgatcg tgatctttct ggagttcttt gcatggggac tgctgaccgc
      841 acctacactg gtggtgctgc acgagacctt tccaaagcac acattcctga tgaatggcct
      901 gatccagggc gtgaagggcc tgctgtcctt tctgtctgcc ccactgatcg gcgccctgtc
      961 cgacgtgtgg ggaaggaagt ctttcctgct gctgaccgtg ttctttacat gcgccccaat
     1021 ccccctgatg aagatctctc cctggtggta tttcgccgtg atcagcgtgt ccggcgtgtt
     1081 cgccgtgacc tttagcgtgg tgttcgccta cgtggccgat atcacacagg agcacgagag
     1141 gagcatggcc tatggcctgg tgtccgccac ctttgccgcc tctctggtga caagccctgc
     1201 aatcggagca tacctgggac gcgtgtatgg cgactccctg gtggtggtgc tggcaaccgc
     1261 aatcgccctg ctggatatct gcttcatcct ggtggccgtg cctgagtctc tgccagagaa
     1321 gatgaggcca gcatcttggg gagcaccaat cagctgggag caggcagacc cctttgcctc
     1381 tctgaagaag gtcggccagg atagcatcgt gctgctgatc tgtatcacag tgttcctgtc
     1441 ctacctgccc gaggccggcc agtacagctc cttctttctg tatctgcggc agatcatgaa
     1501 gtttagccct gagtccgtgg cagccttcat cgccgtgctg ggcatcctga gcatcatcgc
     1561 ccagaccatc gtgctgagcc tgctgatgag atccatcggc aacaagaata caatcctgct
     1621 gggcctgggc tttcagatcc tgcagctggc ctggtatggc ttcggcagcg agccatggat
     1681 gatgtgggca gcaggagcag tggcagcaat gtctagcatc acctttccag ccgtgtccgc
     1741 cctggtgtct aggacagcag acgcagatca gcagggagtg gtgcagggca tgatcaccgg
     1801 catcagagga ctgtgcaacg gactgggacc tgccctgtac ggcttcatct tttatatctt
     1861 tcacgtggag ctgaaggagc tgccaatcac cggcacagac ctgggcacca acacatcccc
     1921 ccagcaccac ttcgagcaga attctatcat ccctggcccc cctttcctgt ttggagcatg
     1981 cagcgtgctg ctggccctgc tggtggccct gttcatcccc gagcacacca atctgtccct
     2041 gaggtcctct agctggcgca agcactgtgg ctctcacagc caccctcaca acacccaggc
     2101 accaggagag gcaaaggagc cactgctgca ggatacaaat gtgtaagacc cagctttctt
     2161 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2221 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2281 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2341 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     2401 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2461 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2521 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2581 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2641 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2701 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2761 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2821 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2881 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2941 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3001 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3061 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3121 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3181 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3241 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3301 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3481 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4021 gtt
//
