LOCUS       sequence-261796-        3492 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 3492)
  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 3492)
  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_SLC25A44
FEATURES             Location/Qualifiers
     source          1..3492
                     /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(1617..1716)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1733..1752)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1734..1752)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1757..1773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1757..1773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1875..1894)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1886..2695
                     /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(1953..1972)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2842..3430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3331..3350
                     /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 accatggagg acaagaggaa catccagatc atcgagtggg agcacctgga
      721 taagaagaag ttctacgtgt tcggcgtggc catgaccatg atgatccggg tgagcgtgta
      781 ccctttcacc ctgatccgga caagactgca ggtgcagaag ggcaagagcc tgtaccacgg
      841 caccttcgac gcctttatca agatcctgag ggccgatggc atcacaggcc tgtatcgcgg
      901 cttcctggtg aacaccttta cactgatcag cggccagtgc tacgtgacca catatgagct
      961 gacccggaag tttgtggccg actacagcca gtccaataca gtgaagtccc tggtggcagg
     1021 aggatctgcc agcctggtgg cccagtccat caccgtgcca atcgatgtgg tgtctcagca
     1081 cctgatgatg cagcggaagg gcgagaagat gggccggttc caggtgagag gaaacccaga
     1141 gggacaggga gtggtggcct ttggccagac caaggacatc atcaggcaga tcctgcaggc
     1201 agatggactg aggggcttct acaggggcta tgtggcctct ctgctgacat acatcccaaa
     1261 tagcgccgtg tggtggccct tctaccactt ttatgccgag cagctgtctt atctgtgccc
     1321 caaggagtgt cctcacatcg tgttccaggc cgtgagcgga cctctggcag cagccaccgc
     1381 ctccatcctg acaaacccaa tggacgtgat caggacccgc gtgcaggtgg agggcaagaa
     1441 ttccatcatc ctgacatttc ggcagctgat ggcagaggag ggaccatggg gactgatgaa
     1501 gggactgtct gccagaatca tctccgccac cccttctaca atcgtgatcg tggtgggcta
     1561 tgagagcctg aagaagctgt ccctgcggcc cgagctggtg gattccagac actgggaccc
     1621 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1681 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     1741 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     1801 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     1861 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     1921 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     1981 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2041 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2101 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2161 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2221 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2281 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2341 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2401 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2461 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2521 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2581 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2641 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2701 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     2761 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     2821 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2881 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2941 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3001 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3061 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3481 tgctcacatg tt
//
