LOCUS       sequence-479354-        3558 bp    DNA     circular SYN 03-AUG-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 3558)
  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 3558)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 3, 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-MAGT1_STOP
FEATURES             Location/Qualifiers
     source          1..3558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..50)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(204..792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(284..303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(939..1748)
                     /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     1662..1681
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1740..1759
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1861..1877
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1861..1877
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1882..1901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1882..1900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1918..2017
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3032..3131)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3147..3164)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3147..3163)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3156..3178)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3227..3313
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3405..3432
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
       61 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac
      841 gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta
      901 atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
      961 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     1021 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     1081 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     1141 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     1201 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     1261 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     1321 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     1381 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     1441 gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     1501 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     1561 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     1621 ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     1681 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata
     1741 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     1801 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc
     1861 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc tggatggcaa
     1921 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
     1981 tttcttataa tgccaacttt gtacaagaaa gctgggtctc agctcatcag aaaagaataa
     2041 gggtagccgt gatacttgga gcggaagatg ctcagcatcc aagaaaagaa cagcaccacc
     2101 agtccgattc ctgccacgca catgatcttc ctcttgccga tatccatgtc agaggtggct
     2161 gcctcgcaca gcagcaccat tcccagtgtc acgccgccat taaacagcag cacgatgtgg
     2221 gtctcggcca cgaactgggc ctggctagag ccgtggatat aattcacgtg gccggtgtgg
     2281 gggttcttgt gggcgtatgg agggccccgg atgtggttcc acatctggcc ggaggtcatg
     2341 gccagcacaa agcacagggc ggcgaaggcc cagcctgtct tattaaacag gaactccatg
     2401 ttgctgcgcc tcagatacac caggcctccg atcacggcca gcagcagtcc cagcatcagt
     2461 ggtcctgcgt aattgggtgg gcggatcact ctgatgttca catcggtgcg gtctgcgatc
     2521 caccttgcga tctgctcggc ggagaagccg cgcacctgca gctcgtaggt atcgcccctc
     2581 ttaggcttgc ccttggcggg aaaattgatg aatgtagggg cagaattcat gttcagcatc
     2641 tgaaacacgt cggagccctc atcgaagtcc accatggcaa agaagattct atttgtaaag
     2701 gcggagctat accgccagct gttggccagg atctggaact cctcatcggc ctgcttgcac
     2761 accacgcact gtctgtgcag ctgcagggcg gtaaacatca cgatcacaga gtagttccga
     2821 gggggggcct tcaccagtct ccggaacttg tcgccattca ttctgatcac gggccgcttg
     2881 tttgtccact ccatcagctg ggacaccttc tcgctcagca ccatctcctt cttccgctgg
     2941 gcagaggcgg atggcacgtc gcacacgatc agcagggcca ccaccattgt cacggacacg
     3001 caccagaatc tccaccgggc ggccatggtg gagcctgctt ttttgtacaa agttggcatt
     3061 ataaaaaagc attgctcatc aatgtgttgc aacgaacagg tcactatcag tcaaaataaa
     3121 atcattattt ggggcccgag cttaagactg gccgtcgttt tacaacgtcg tgactgggaa
     3181 aacatccatg ctagcgttaa cgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     3241 ggctcagtcg gaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     3301 gagtaggaca aatccgccgg gagcggattt gaacgttgtg aagcaacggc ccggagggtg
     3361 gcgggcagga cgcccgccat aaactgccag gcatcaaact aagcagaagg ccatcctgac
     3421 ggatggcctt tttgcgtttc tacaaactct tcctggctag cggtacgcgt attaattgcg
     3481 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3541 ggccaacgcg cggggaga
//
