LOCUS       sequence-479355-        3441 bp    DNA     circular SYN 14-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 3441)
  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 3441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 14, 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-TMEM241_STOP
FEATURES             Location/Qualifiers
     source          1..3441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(861..1670)
                     /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     1584..1603
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1662..1681
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1783..1799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1783..1799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1804..1823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1804..1822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1840..1939
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2837..2936)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2952..2969)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2952..2968)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2961..2983)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3032..3118
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3210..3237
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3396..3413)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact cacgttaagg
      781 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg ccagtgttac
      841 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      901 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      961 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1021 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1081 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1141 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1201 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     1261 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1321 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca
     1381 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1441 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1501 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     1561 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1621 atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt
     1681 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     1741 gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac agctatgacc
     1801 atgtaatacg actcactata ggggatatca gctggatggc aaataatgat tttattttga
     1861 ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat aatgccaact
     1921 ttgtacaaga aagctgggtc tcaggagctc ttcctctcgg aaaacaccag cagggcctct
     1981 cccagggcgc ccagcagcag acatcctgtg gtggcgcttg tcaggatggc gtcgaacagc
     2041 aggatagaca ggccggcggt gatgatcttg gcaaagaaga tccatgctgc gcactgtcct
     2101 ggtgccagca gattcttcag cttcactgtg ctaaacatca gaaagaagcc caggaagccg
     2161 gatgcacagc agcttccgtg aaaccgatag aagtacagaa aggggaaatc cagcacgcta
     2221 aacaggtcgc cggttgggtg agaggcaaag gccagcagca ccacggagaa gatatagttc
     2281 aggtactgct gatcgatgtc gctcagggca gaaggcttct gagacttctg caggatctta
     2341 taggcgccca cgcacagcag gtggatgatg gcccagaagt agccatcagg attaaactgg
     2401 gagtcgttga agggcagaca gcctgctgct gccagcagca gcagggcgct gcagatcttg
     2461 gcgggagatg tcttctcctt ctgaaaacac ttctgatagc cgcagatgat cacttcggcc
     2521 acgttgtgca gggtcaggaa cactgggatt gccagcctag acagggccct ggatcctgcg
     2581 tagatgatgc ccacgaacag cacggatgca ggcagccaca ccagcacgtg gctccgagag
     2641 gagctattga tctccaccca gcccagcttc caggacacgt gcagcagcag gccgccgatc
     2701 agtgtctgcc agccctgaaa cagtgttgga taggtgaact tcagcacaga cagcacgtac
     2761 ttgttggtca gataggaggc caggtaacat gtgcagaagg tcaggcccac caggcttctc
     2821 cgcacgcaca tggtggagcc tgcttttttg tacaaagttg gcattataaa aaagcattgc
     2881 tcatcaatgt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttggggc
     2941 ccgagcttaa gactggccgt cgttttacaa cgtcgtgact gggaaaacat ccatgctagc
     3001 gttaacgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga
     3061 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3121 gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc
     3181 gccataaact gccaggcatc aaactaagca gaaggccatc ctgacggatg gcctttttgc
     3241 gtttctacaa actcttcctg gctagcggta cgcgtattaa ttgcgttgcg ctcactgccc
     3301 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3361 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3421 gtcgttcggc tgcggcgagc g
//
