LOCUS       sequence-260257-        3444 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 3444)
  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 3444)
  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_SLC25A4
FEATURES             Location/Qualifiers
     source          1..3444
                     /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(1569..1668)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1685..1704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1686..1704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1709..1725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1709..1725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1827..1846)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1838..2647
                     /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(1905..1924)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2794..3382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3283..3302
                     /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 accatgggcg accacgcctg gtccttcctg aaggattttc tggcaggagg
      721 agtggcagca gccgtgtcta agaccgccgt ggcccccatc gagagagtga agctgctgct
      781 gcaggtgcag cacgccagca agcagatctc cgccgagaag cagtataagg gcatcatcga
      841 ttgcgtggtg aggatcccaa aggagcaggg cttcctgagc ttttggcgcg gcaacctggc
      901 caatgtcata cgctacttcc ccacacaggc cctgaacttc gcctttaagg ataagtacaa
      961 gcagctgttt ctgggcggcg tggacagaca caagcagttc tggaggtatt ttgccggcaa
     1021 tctggcatcc ggaggagcag caggagccac ctctctgtgc ttcgtgtacc ctctggattt
     1081 tgcaaggaca aggctggcag cagacgtggg caagggcgcc gcccagcgcg agttccacgg
     1141 cctgggcgat tgtatcatca agatctttaa gagcgacgga ctgaggggac tgtaccaggg
     1201 cttcaacgtg tccgtgcagg gcatcatcat ctaccgggcc gcctattttg gcgtgtacga
     1261 caccgccaag ggcatgctgc ccgatcctaa gaatgtgcac atcttcgtgt cttggatgat
     1321 cgcacagagc gtgaccgcag tggcaggact ggtgtcttat ccatttgata cagtgcggag
     1381 aaggatgatg atgcagagcg gcagaaaggg cgccgacatc atgtacaccg gcacagtgga
     1441 ttgttggagg aagatcgcca aggacgaggg cgccaaggcc ttctttaagg gcgcctggtc
     1501 caacgtgctg aggggaatgg gaggagcctt cgtgctggtg ctgtatgacg agatcaagaa
     1561 gtacgtggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1621 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1681 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1741 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1801 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1861 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1921 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     1981 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2041 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2101 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2161 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2221 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2281 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2341 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2401 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2461 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2521 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2581 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2641 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2701 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2941 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3421 ttgctggcct tttgctcaca tgtt
//
