LOCUS       sequence-479317-        4074 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 4074)
  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 4074)
  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-SLC41A3_STOP
FEATURES             Location/Qualifiers
     source          1..4074
                     /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(3548..3647)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3663..3680)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3663..3679)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3672..3694)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3743..3829
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3921..3948
                     /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 gctgggtctt agatgcttgt cttggccagc
     2041 agacagaagg gaaaatagaa agacagcagg gacacgatca ggtgctgggt acagatgaag
     2101 ggctggatca gtccccatcc tggagatgct gtgcagcgcc taggcactgc tgcggtgtgt
     2161 cccacggagc tagatcccag cagatcgccc agtccggtca ggtaggggat gcagtgatta
     2221 tctgggtcca gggcctggtg ccatgtcagc cgcaccatca cttcggccag atacagcagg
     2281 atggtcacct ggatcaggcc ggccagcagg tacagcacca caaaggtctg gctgttgatc
     2341 acagactggc cctccaccag atagatgatg taaaagaaga tcaggtggcc tggcaccacc
     2401 agcagcagca gcactctggc agacatggaa ttgatctcag atgtacagaa ggtggagcag
     2461 gggtttggcc aaaacttctt catctgcagt ggcagcacgc caggggcgct ccacatgtgc
     2521 agatatgtgc tgatccgaga ggtctggatg gccaccagat tgccgcccac gccacagatc
     2581 actggtgtga agatggccat gcccttgtac tgctgcttgg acacggtctt gctcaggatc
     2641 aggccgccaa aggagctgat gaccatggcc aggatgatag ggaaccagcc aaacttcagg
     2701 atcttcacga tgggtgggga ctgcttggcg atcagcaccc acacgggtgt cagggcggcg
     2761 aaggacaggc acaccagtgg ggtcagatat ctgctatcct tgtgccggta aaagaaagag
     2821 gacaccaggg ccaggataga cagtgtgatc agatcgccca gggatgctgc gataggggtt
     2881 gcgatattgt ctgggttcac tcccagcttc cttgctccga tcacgatgca caccatcagc
     2941 acgcccaggg cgaaggctgc cagaaaggct gtcagcacgc tagaggcgca cagcagctcc
     3001 accttggcca cgtccacctc ctccctggac accacgccca gcagcagggc ggccactgct
     3061 gccagcagtc ccaccacggt ggcctgcacc tggatcaggg ccaggttgga gctgatcact
     3121 ctgtgctgct cctgaggatc gtcgatctgt cctgtatttg ctgcggtgga cagcctgctt
     3181 gccagtgtca tctccaggtt gcccttcagg cccaccagag gtggcaccag ggtcagcaga
     3241 tccttcacct ccacgaacac gggccagtgc tgaaagtagt ccagcagcat tcctgcccag
     3301 ctcagtccca gtcctgcaaa catgaagggc acggtcacct gcaggccgat ggaccatgtg
     3361 gtctcgcggc ttggctctgt ctccagaggc ttgggggtca cgctctgaga ctctggtgcc
     3421 ctcagggcgc cgtcctcgct tgccacaggc agtcctccgg tagacagtgg gtgaggcagt
     3481 cccagctctc caggcttgcc gcaggaatcc agtctccgct gccttgtctc ggtgccgtcc
     3541 atggtggagc ctgctttttt gtacaaagtt ggcattataa aaaagcattg ctcatcaatg
     3601 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgggg cccgagctta
     3661 agactggccg tcgttttaca acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg
     3721 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcggaag actgggcctt
     3781 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     3841 ggatttgaac gttgtgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     3901 tgccaggcat caaactaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     3961 aactcttcct ggctagcggt acgcgtatta attgcgttgc gctcactgcc cgctttccag
     4021 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gaga
//
