LOCUS       sequence-262986-        3522 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 3522)
  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 3522)
  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-TMEM165
FEATURES             Location/Qualifiers
     source          1..3522
                     /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(1647..1746)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1763..1782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1764..1782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1787..1803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1787..1803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1905..1924)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1916..2725
                     /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(1983..2002)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2872..3460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3361..3380
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggcag cagcagcacc tggaaacgga agggcatctg ccccaagact
      721 gctgctgctg ttcctggtgc cactgctgtg ggcaccagca gccgtgcggg caggaccaga
      781 cgaggatctg agccacagaa ataaggagcc acctgcacca gcacagcagc tgcagccaca
      841 gcccgtggcc gtgcagggcc ctgagccagc ccgggtggag aagatcttta cccccgccgc
      901 ccctgtgcac acaaacaagg aggacccagc cacccagaca aatctgggct tcatccacgc
      961 ctttgtggcc gccatctctg tgatcatcgt gagcgagctg ggcgataaga ccttctttat
     1021 cgccgccatc atggccatgc ggtacaacag actgaccgtg ctggcaggag caatgctggc
     1081 cctgggcctg atgacatgcc tgtctgtgct gttcggctac gccaccacag tgatccctcg
     1141 ggtgtatacc tactatgtga gcacagtgct gttcgccatc tttggcatca ggatgctgcg
     1201 cgagggcctg aagatgtccc cagacgaggg acaggaggag ctggaggagg tgcaggccga
     1261 gctgaagaag aaggacgagg agttccagag aaccaagctg ctgaatggac caggcgatgt
     1321 ggagaccggc acatccatca cagtgcccca gaagaagtgg ctgcacttca tctctcctat
     1381 ctttgtgcag gccctgaccc tgacatttct ggcagagtgg ggcgacaggt cccagctgac
     1441 cacaatcgtg ctggcagcaa gggaggatcc ttatggagtg gcagtgggag gaaccgtggg
     1501 acactgcctg tgcacaggac tggccgtgat cggaggaagg atgatcgccc agaagatcag
     1561 cgtgcgcacc gtgacaatca tcggcggcat cgtgtttctg gccttcgcct tttccgccct
     1621 gttcatcagc cctgattccg gctttgaccc agctttcttg tacaaagttg gcattataag
     1681 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1741 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1801 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1861 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1921 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1981 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2041 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2101 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2161 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2221 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2281 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2341 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2401 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2461 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2521 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2581 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2641 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2701 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2761 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2821 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2881 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2941 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3001 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3061 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3121 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3181 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3241 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3301 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3361 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3421 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3481 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
