LOCUS       sequence-421010-        4587 bp    DNA     circular SYN 14-APR-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 4587)
  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 4587)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC25A12_STOP
FEATURES             Location/Qualifiers
     source          1..4587
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(82..181)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(198..217)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(199..217)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(222..238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(222..238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(340..359)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             351..1160
                     /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(418..437)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1307..1895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1796..1815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2049..2066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2225..2252)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2344..2430)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2479..2501
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2493..2510
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2494..2510
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2526..2625
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3502..3516
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
ORIGIN
        1 tcggcctgta tctgcctaag tttaagtccc catctgtggc cgtggtgcag cccaaggccg
       61 ccgtggccgc cacccagtaa gacccagctt tcttgtacaa agttggcatt ataagaaagc
      121 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
      181 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
      241 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
      301 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      361 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
      421 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
      481 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
      541 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
      601 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
      661 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
      721 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
      781 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
      841 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
      901 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
      961 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     1021 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     1081 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     1141 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     1201 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     1261 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1321 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1381 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1441 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1501 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1561 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1621 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1681 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1741 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1801 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1861 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1921 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1981 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2041 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2101 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2161 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     2221 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     2281 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     2341 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     2401 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     2461 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
     2521 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa cacattgatg
     2581 agcaatgctt ttttataatg ccaactttgt acaaaaaagc aggctccacc atggccgtga
     2641 aggtgcagac cacaaagagg ggcgacccac acgagctgcg caacatcttc ctgcagtacg
     2701 ccagcaccga ggtggacggc gagaggtata tgacacccga ggattttgtg cagcggtacc
     2761 tgggcctgta taatgatccc aactccaatc ctaagatcgt gcagctgctg gcaggagtgg
     2821 cagaccagac caaggatggc ctgatctctt accaggagtt cctggccttt gagtccgtgc
     2881 tgtgcgcccc tgactctatg ttcatcgtgg ccttccagct gtttgataag tccggcaacg
     2941 gcgaggtgac ctttgagaat gtgaaggaga tcttcggcca gacaatcatc caccaccaca
     3001 tccccttcaa ctgggactgt gagtttatcc ggctgcactt cggccacaac agaaagaagc
     3061 acctgaatta taccgagttc acacagtttc tgcaggagct gcagctggag cacgcaaggc
     3121 aggccttcgc cctgaaggat aagtctaaga gcggcatgat cagcggcctg gacttctccg
     3181 atatcatggt gaccatcaga tctcacatgc tgacaccctt tgtggaggag aatctggtga
     3241 gcgccgcagg aggatccatc tctcaccagg tgagcttctc ctactttaac gccttcaatt
     3301 ccctgctgaa caatatggag ctggtgcgga agatctattc taccctggcc ggcacaagaa
     3361 aggacgtgga ggtgaccaag gaggagttcg cacagagcgc catcagatac ggacaggtga
     3421 cacctctgga gatcgacatc ctgtaccagc tggccgatct gtataacgcc tccggcaggc
     3481 tgaccctggc agatatcgag aggatcgcac cactggcaga gggcgccctg ccttataatc
     3541 tggcagagct gcagaggcag cagtccccag gactgggcag acccatctgg ctgcagatcg
     3601 cagagagcgc ctaccggttc accctgggct ccgtggcagg agcagtggga gcaaccgccg
     3661 tgtatcccat cgacctggtg aagacaagga tgcagaacca gaggggatct ggaagcgtgg
     3721 tgggagagct gatgtacaag aattcttttg attgcttcaa gaaggtgctg cggtacgagg
     3781 gcttctttgg cctgtataga ggcctgatcc cacagctgat cggagtggca ccagagaagg
     3841 ccatcaagct gaccgtgaac gactttgtga gggataagtt cacacggaga gacggcagcg
     3901 tgccactgcc agcagaggtg ctggcaggag gctgtgccgg cggctcccag gtcatcttca
     3961 ccaaccctct ggagatcgtg aagatcaggc tgcaggtggc aggagagatc accacaggac
     4021 caagggtgtc tgccctgaat gtgctgcgcg acctgggcat cttcggcctg tacaagggcg
     4081 ccaaggcctg ctttctgcgc gatatccctt ttagcgccat ctacttccca gtgtatgccc
     4141 actgtaagct gctgctggca gatgagaacg gacacgtggg aggcctgaat ctgctggcag
     4201 caggagcaat ggcaggagtg ccagccgcca gcctggtgac ccctgccgac gtgatcaaga
     4261 caaggctgca ggtggcagca agagcaggac agaccacata ctctggcgtg atcgattgct
     4321 tccggaagat cctgagagag gagggcccaa gcgccttttg gaagggaacc gcagcaaggg
     4381 tgtttcgcag ctccccacag ttcggagtga ccctggtgac atacgagctg ctgcagcggt
     4441 ggttttatat cgacttcgga ggcctgaagc cagcaggatc tgagcctaca ccaaagagca
     4501 ggatcgcaga cctgccacct gcaaaccctg atcacatcgg aggatacagg ctggcaaccg
     4561 ccacatttgc cggcatcgag aataagt
//
