LOCUS       sequence-430411-        3450 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 3450)
  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 3450)
  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-SLC25A21_STOP
FEATURES             Location/Qualifiers
     source          1..3450
                     /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(1007..1106)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1123..1142)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1124..1142)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1147..1163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1147..1163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1265..1284)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1276..2085
                     /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(1343..1362)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2232..2820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2721..2740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2974..2991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3150..3177)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3269..3355)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3404..3426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3418..3435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3419..3435
                     /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 ccaccatgag cgccaagcca
      121 gaggtgtccc tggtgcggga ggcctctaga cagatcgtgg caggaggatc tgccggactg
      181 gtggagatct gccttatgca cccgttggat gtcgtaaaga ccaggttcca gatccagcgg
      241 tgcgccacag accctaactc ttataagagc ctggtggata gcttccggat gatctttcag
      301 atggagggcc tgttcggctt ttacaagggc atcctgcccc ctatcctggc cgagacccct
      361 aagagagccg tgaagttctt tacattcgag cagtataaga agctgctggg ctacgtgagc
      421 ctgtccccag ccctgacctt tgccatcgca ggactgggaa gcggactgac agaggccatc
      481 gtggtgaatc cattcgaggt ggtgaaagtg ggcctgcagg ccaaccggaa tacctttgcc
      541 gagcagccct ccacagtggg ctatgccaga cagatcatca agaaggaggg atggggactg
      601 cagggcctga acaagggact gaccgccaca ctgggcaggc acggcgtgtt taatatggtg
      661 tacttcggct tttactataa cgtgaagaat atgatcccag tgaacaagga ccccatcctg
      721 gagttctggc gcaagtttgg aatcggactg ctgtccggaa ccatcgcatc tgtgatcaat
      781 atccctttcg atgtggccaa gtccaggatc cagggaccac agccagtgcc tggagagatc
      841 aagtatcgca catgctttaa gaccatggcc acagtgtatc aggaggaggg catcctggcc
      901 ctgtacaagg gcctgctgcc taagatcatg aggctgggac caggaggagc cgtgatgctg
      961 ctggtgtacg agtataccta ctcttggctg caggagaact ggtaagaccc agctttcttg
     1021 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1081 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     1141 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     1201 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     1261 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     1321 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     1381 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     1441 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     1501 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     1561 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     1621 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     1681 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     1741 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     1801 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     1861 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     1921 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     1981 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     2041 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     2101 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     2161 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     2221 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2281 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2341 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2401 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2461 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2521 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2581 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2641 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2701 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2761 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2821 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2881 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2941 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3001 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3061 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     3121 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     3181 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     3241 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     3301 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     3361 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     3421 aaaacgacgg ccagtcttaa gctcgggccc
//
