LOCUS       sequence-338289-        4029 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 4029)
  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 4029)
  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-SVOPL_STOP
FEATURES             Location/Qualifiers
     source          1..4029
                     /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"
     regulatory      1433..1442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1586..1685)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1702..1721)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1703..1721)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1726..1742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1726..1742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1844..1863)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1855..2664
                     /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(1922..1941)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2811..3399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3300..3319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3553..3570
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3729..3756)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3848..3934)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3983..4005
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3997..4014
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3998..4014
                     /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 ccaccatggc caccaagccc
      121 acagagcctg tgaccatcct gtctctgagg aagctgagcc tgggcacagc cgagccacag
      181 gtgaaggagc ccaagacctt tacagtggag gacgcagtgg agaccatcgg attcggccgc
      241 tttcacatcg ccctgttcct gatcatgggc tccacaggcg tggtggaggc catggagatc
      301 atgctgatcg ccgtggtgtc tccagtgatc aggtgcgagt ggcagctgga gaactggcag
      361 gtggccctgg tgaccacaat ggtgttcttt ggctacatgg tgttcagcat cctgtttggc
      421 ctgctggccg atcggtatgg cagatggaag atcctgctga tctccttcct gtggggcgcc
      481 tacttttctc tgctgaccag cttcgccccc tcctatatct ggttcgtgtt tctgaggaca
      541 atggtgggat gtggcgtgag cggacactcc cagggactga tcatcaagac cgagtttctg
      601 ccaacaaagt accgcggcta tatgctgcca ctgagccagg tgttctggct ggcaggatcc
      661 ctgctgatca tcggactggc ctctgtgatc atcccgacta taggatggcg atggttgatc
      721 agggtggcat ccatccctgg catcatcctg atcgtggcct tcaagtttat cccagagtcc
      781 gccaggttca acgtgtctac cggaaataca agggccgccc tggccaccct ggagagggtg
      841 gccaagatga atagaagcgt gatgcctgag ggcaagctgg tggagccagt gctggagaag
      901 aggggccgct ttgccgacct gctggatgcc aagtacctgc ggaccacact gcagatctgg
      961 gtcatctggc tgggcatctc tttcgcctac tatggcgtga tcctggcaag cgccgagctg
     1021 ctggagagag acctggtgtg cggctccaag tctgatagcg ccgtggtggt gaccggaggc
     1081 gactccggcg agtcccagtc tccctgctac tgtcacatgt ttgcccctag cgattatcgg
     1141 accatgatca tctccacaat cggcgagatc gccctgaacc cactgaatat cctgggcatc
     1201 aacttcctgg gccggagact gagcctgtcc atcaccatgg gctgcacagc cctgttcttt
     1261 ctgctgctga atatctgtac cagctccgcc ggcctgatcg gcttcctgtt tatgctgaga
     1321 gccctggtgg ccgccaactt taataccgtg tacatctata cagccgaggt gtatccaacc
     1381 acaatgcggg ccctgggaat gggcacatct ggaagcctgt gcagaattgg cgccatggtg
     1441 gcccctttca tctctcaggt gctgatgtcc gcctctatcc tgggcgccct gtgcctgttt
     1501 tctagcgtgt gcgtggtgtg cgccatcagc gccttcaccc tgccaatcga gacaaaggga
     1561 cgggccctgc agcagatcaa gtaagaccca gctttcttgt acaaagttgg cattataaga
     1621 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1681 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1741 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1801 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1861 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1921 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     1981 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2041 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2101 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2161 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2221 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2281 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2341 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2401 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2461 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2521 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2581 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2641 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2701 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2761 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3481 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3541 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3601 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3661 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     3721 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     3781 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     3841 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3901 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     3961 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     4021 ctcgggccc
//
