LOCUS       sequence-384582-        3633 bp    DNA     circular SYN 15-DEC-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 3633)
  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 3633)
  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-LETMD1_STOP
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1190..1289)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1306..1325)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1307..1325)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1330..1346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1330..1346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1448..1467)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1459..2268
                     /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(1526..1545)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2415..3003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2904..2923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3157..3174
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3333..3360)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3452..3538)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3587..3609
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3601..3618
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3602..3618
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cctgtccaga
      121 gtgtgctggg caaggtccgc cgtgtgggga tctgccgtga cccctggcca ctttgtgaca
      181 cggagactgc agctgggcag atctggactg gcatggggag caccaaggag ctccaagctg
      241 cacctgagcc ccaaggccga cgtgaagaac ctgatgtcct acgtggtgac caagacaaag
      301 gccatcaatg gcaagtatca caggtttctg ggccgccact tccctcggtt ttacgtgctg
      361 tataccatct tcatgaaggg cctgcagatg ctgtgggcag acgccaagaa ggcaaggcgc
      421 atcaagacaa acatgtggaa gcacaatatc aagtttcacc agctgccata ccgcgagatg
      481 gagcacctgc ggcagtttag acaggatgtg accaagtgtc tgttcctggg catcatcagc
      541 atcccccctt tcgccaacta cctggtgttt ctgctgatgt atctgttccc tcgccagctg
      601 ctgatccggc acttctggac cccaaagcag cagacagact ttctggatat ctaccacgcc
      661 ttcagaaagc agagccaccc cgagatcatc tcctatctgg agaaagtgat ccctctgatc
      721 agcgacgcag gactgaggtg gaggctgacc gatctgtgca caaagatcca gaggggaacc
      781 cacccagcaa tccacgatat cctggccctg cgggagtgtt tctccaacca ccccctgggc
      841 atgaatcagc tgcaggccct gcacgtgaag gccctgtcta gagccatgct gctgaccagc
      901 tacctgccac cacctctgct gaggcacaga ctgaagacac acaccacagt gatccaccag
      961 ctggacaagg ccctggcaaa gctgggaatc ggacagctga cagcccagga ggtgaagtcc
     1021 gcctgctatc tgaggggcct gaattctacc cacatcggcg aggataggtg caggacatgg
     1081 ctgggagagt ggctgcagat ctcttgtagc ctgaaggagg ccgagctgtc tctgctgctg
     1141 cacaacgtgg tgctgctgag caccaattat ctgggcacac ggagatgaga cccagctttc
     1201 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1261 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     1321 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1381 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1441 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1501 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1561 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1621 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1681 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1741 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1801 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1861 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1921 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1981 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     2041 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2101 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2161 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2221 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2281 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2341 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2521 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3121 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3181 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3241 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3301 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3361 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3421 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3481 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3541 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3601 tgtaaaacga cggccagtct taagctcggg ccc
//
