LOCUS       sequence-312250-        3960 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 3960)
  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 3960)
  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-SLC25A25_STOP
FEATURES             Location/Qualifiers
     source          1..3960
                     /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(2085..2184)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2201..2220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2202..2220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2225..2241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2225..2241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2343..2362)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2354..3163
                     /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(2421..2440)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3310..3898
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3799..3818
                     /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 caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatgctgt gcctgtgcct gtacgtgcca gtgatcggcg aggcccagac
      721 agagttccag tattttgagt ccaagggcct gcccgccgag ctgaagagca tcttcaagct
      781 gtccgtgttt atcccttccc aggagttttc tacctaccgg cagtggaagc agaagatcgt
      841 gcaggccggc gacaaggatc tggacggcca gctggacttc gaggagtttg tgcactatct
      901 gcaggatcac gagaagaagc tgagactggt gttcaagtcc ctggataaga agaacgacgg
      961 ccggatcgat gcccaggaga tcatgcagtc tctgagagac ctgggcgtga agatcagcga
     1021 gcagcaggcc gagaagatcc tgaagtctat ggataagaac ggcaccatga caatcgactg
     1081 gaatgagtgg cgggattacc acctgctgca ccccgtggag aacatccctg agatcatcct
     1141 gtattggaag cacagcacaa tctttgacgt gggcgagaat ctgaccgtgc ctgatgagtt
     1201 cacagtggag gagaggcaga ccggaatgtg gtggaggcac ctggtggcag gaggaggagc
     1261 aggagccgtg agccggacct gcacagcccc actggacaga ctgaaggtgc tgatgcaggt
     1321 gcacgccagc aggtccaaca atatgggcat cgtgggcggc tttacacaga tgatcaggga
     1381 gggaggagca cggtctctgt ggagaggcaa cggcatcaat gtgctgaaga tcgcccccga
     1441 gagcgccatc aagttcatgg cctacgagca gatcaagagg ctggtgggct ccgaccagga
     1501 gaccctgagg atccacgagc gcctggtggc aggatctctg gcaggagcaa tcgcacagag
     1561 ctccatctac cctatggagg tgctgaagac acggatggcc ctgagaaaga ccggccagta
     1621 ttccggcatg ctggattgtg caaggagaat cctggcaagg gagggagtgg ccgcctttta
     1681 caagggctat gtgccaaata tgctgggcat catcccctac gccggcatcg acctggccgt
     1741 gtatgagaca ctgaagaacg cctggctgca gcactacgcc gtgaattctg ccgatcctgg
     1801 cgtgttcgtg ctgctggcat gcggaaccat gtctagcaca tgtggccagc tggccagcta
     1861 tccactggcc ctggtgagga ccaggatgca ggcacaggca tccatcgagg gagcacccga
     1921 ggtgacaatg tcctctctgt tcaagcacat cctgaggacc gagggagcct ttggcctgta
     1981 caggggactg gcccctaact tcatgaaagt gatcccagcc gtgtctatca gctacgtggt
     2041 gtatgagaat ctgaagatca ccctgggcgt gcagtccaga taagacccag ctttcttgta
     2101 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2161 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2221 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2281 tgcacaagat aaaataatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2341 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2401 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2461 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2521 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2581 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2641 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2701 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2761 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2821 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2881 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2941 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3001 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3061 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3121 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3181 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3241 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3301 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3361 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3421 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3481 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3541 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3601 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3661 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3721 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3781 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3841 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3901 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
