LOCUS       sequence-258979-        3864 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 3864)
  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 3864)
  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_CLN3
FEATURES             Location/Qualifiers
     source          1..3864
                     /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=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1989..2088)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2105..2124)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2106..2124)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2129..2145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2129..2145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2247..2266)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2258..3067
                     /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(2325..2344)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3214..3802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3703..3722
                     /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 caacaaattg ataagcaatg ctttcttata atgccaactt tgtacaagaa
      661 agctgggtcg ctcagctggc acaggaagtc gtgcagaggc agggccagca gtccggacag
      721 gctgattccc agtgtatcgg agatgcaggt ggcggccatg gcaaactccc ggtgctcgtc
      781 gcttgtctcc agggcgatat tgtggaaggt gttcacatat gctgctcctc ccagcagtcc
      841 ctcatacagg atgatcagaa acaccaggta gatgctgggc agaaagccga accacacgtc
      901 ggccagcagg aacaccaggt tcagacactg cagcagggcc agggcccatg taaacctgat
      961 tctacagcac cgcaggctag agcgagaggc gaacacgccg gcctgataca gcatctgata
     1021 ccagcggtac tgctgggcgt gagacaggga ggtattccaa aagaacagca gctcaaacag
     1081 gccctggttg atgaagtact cggcgaagta caccaccacc agtggcacga tgtaccacag
     1141 caggcccttg aacactgtcc accgctccct caggctcagg gagctagagg atcctggctt
     1201 agactctggg gcctcggtcc tgatcagagg ctgtcttgcg gcgctctctg cctcctcctc
     1261 tcctcctggg tcctgggcct caggagaggt cagcagcagg aagtaggatg ccagcagcag
     1321 ggcggggatt cccagcatgc tcagcagtgt ctgctgtgga gacagtcctg cctgggtcag
     1381 tcccagatag gacagggcgc ccagcagtcc tgctcctcct gttccgctag accaccaaga
     1441 gatcacggcc ctagggtaaa aggctgtcag ggacaggaag gtcacctctc ccagtccgga
     1501 gctgatggat gcaaacacca ctccgcacag gctggtgccc acgctgtgag aaaaggccac
     1561 cagcacgaag gatcctgctg cgcagattcc ggacaccagc actctagggc tatagggcag
     1621 caggtgcagt cccagaggtg ccagcagctt gatgaccagg gtgggcagga tatctgccag
     1681 cagcacggct gcggtggaca cgctattgca gtcgaaccga gaggagctgt tgtgtgggat
     1741 aggtgtgggg cctggatcca cgtggctctg attgccagag gtgcgcttgt ggctcaggat
     1801 gtcgtgggcg gcagacagca tgaccacgta ggaaaaattg ttgcacagtc ccagcagcca
     1861 gaatcccact gcgttcttcc agtgtgctcc ctggtgatcc agcagtggca gcctaggctc
     1921 tggcacggtc tcctctccct cagagtcgga gaaccttctc cgggatcctg cgcatcctcc
     1981 catggtggag cctgcttttt tgtacaaagt tggcattata aaaaagcatt gctcatcaat
     2041 gtgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2101 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2161 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2221 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2281 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2341 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2401 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2461 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2521 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2581 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     2641 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     2701 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     2761 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     2821 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     2881 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     2941 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3001 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3061 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3121 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3841 ttgctggcct tttgctcaca tgtt
//
