LOCUS       sequence-260990-        3504 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 3504)
  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 3504)
  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_SLC25A42
FEATURES             Location/Qualifiers
     source          1..3504
                     /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"
     regulatory      956..965
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1629..1728)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1745..1764)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1746..1764)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1769..1785)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1769..1785)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1887..1906)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1898..2707
                     /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(1965..1984)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2854..3442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3343..3362
                     /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 accatgggca acggcgtgaa ggagggaccc gtgcggctgc acgaggacgc
      721 agaggccgtg ctgagctcct ctgtgagctc caagcgcgat caccggcagg tgctgtctag
      781 cctgctgagc ggcgccctgg caggcgccct ggcaaagacc gcagtggcac ctctggacag
      841 aacaaagatc atcttccagg tgtcctctaa gcggttcagc gccaaggagg cctttcgggt
      901 gctgtactat acctacctga acgagggctt cctgtctctg tggagaggca atagcgccac
      961 catggtgagg gtggtgcctt acgccgccat ccagttttct gcccacgagg agtataagcg
     1021 catcctgggc agctactatg gcttccgggg agaggccctg ccaccttggc caagactgtt
     1081 tgcaggcgcc ctggcaggaa ccacagcagc atccctgacc tacccactgg atctggtgag
     1141 agccaggatg gccgtgacac ctaaggagat gtatagcaac atcttccacg tgtttatcag
     1201 aatctccagg gaggagggcc tgaagaccct gtaccacggc tttatgccta cagtgctggg
     1261 cgtgatccca tacgccggcc tgagcttctt tacctatgag acactgaagt ccctgcacag
     1321 ggagtactct ggccggagac agccatatcc cttcgagcgc atgatctttg gagcatgcgc
     1381 aggactgatc ggacagagcg cctcctaccc actggacgtg gtgaggcgcc ggatgcagac
     1441 cgcaggagtg acaggatatc caagggcatc tatcgcacgc accctgagga caatcgtgag
     1501 agaggaggga gccgtgaggg gactgtataa gggcctgtcc atgaattggg tgaagggccc
     1561 catcgccgtg ggcatctctt tcaccacatt tgatctgatg cagatcctgc tgcggcacct
     1621 gcagagcgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1681 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     1741 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     1801 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     1861 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     1921 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1981 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2041 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2101 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2161 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2221 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2281 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2341 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2401 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2461 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2521 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2581 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     2641 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     2701 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     2761 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     2821 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2881 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2941 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3001 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3061 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3121 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3181 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3241 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3301 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3361 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3421 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3481 ttgctggcct tttgctcaca tgtt
//
