LOCUS       sequence-263284-        4134 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 4134)
  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 4134)
  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_SPNS1
FEATURES             Location/Qualifiers
     source          1..4134
                     /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(2259..2358)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2375..2394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2376..2394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2399..2415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2399..2415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2517..2536)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2528..3337
                     /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(2595..2614)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3484..4072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3973..3992
                     /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 accatggccg gctctgacac cgcccctttc ctgagccagg ccgacgatcc
      721 agacgatgga ccagtgccag gaacccctgg actgccaggc tctacaggca accccaagag
      781 cgaggagcca gaggtgcctg accaggaggg actgcagagg atcacaggac tgtcccctgg
      841 ccggtctgcc ctgatcgtgg ccgtgctgtg ctacatcaac ctgctgaatt atatggatcg
      901 ctttaccgtg gcaggcgtgc tgccagacat cgagcagttc tttaacatcg gcgatagctc
      961 ctctggcctg atccagacag tgttcatcag ctcctacatg gtgctggccc ccgtgttcgg
     1021 ctatctgggc gatcgctaca atcggaagta tctgatgtgc ggcggcatcg ccttctggag
     1081 cctggtgacc ctgggctcta gcttcatccc aggcgagcac ttttggctgc tgctgctgac
     1141 aaggggactg gtgggagtgg gagaggcaag ctactccacc atcgcaccta cactgatcgc
     1201 cgacctgttt gtggccgatc agagaagcag gatgctgtcc atcttctact ttgccatccc
     1261 agtgggaagc ggactgggat atatcgcagg atccaaggtg aaggacatgg ccggcgattg
     1321 gcactgggca ctgagagtga cccctggact gggagtggtg gccgtgctgc tgctgttcct
     1381 ggtggtgaga gagccaccta ggggagcagt ggagaggcac tctgacctgc cacccctgaa
     1441 cccaacaagc tggtgggcag atctgcgcgc cctggcaagg aatccatcct tcgtgctgtc
     1501 ctctctgggc ttcaccgccg tggcctttgt gacaggatcc ctggccctgt gggcacctgc
     1561 ctttctgctg cgctctaggg tggtgctggg agagacccct ccatgcctgc caggcgactc
     1621 ctgtagctcc tctgattctc tgatctttgg cctgatcacc tgcctgacag gcgtgctggg
     1681 agtgggactg ggagtggaga tcagccggag actgagacac tccaacccaa gggcagaccc
     1741 actggtgtgc gcaaccggac tgctgggctc cgcccccttc ctgtttctgt ctctggcatg
     1801 tgcaagggga agcatcgtgg caacctacat cttcatcttt atcggcgaga cactgctgag
     1861 catgaattgg gccatcgtgg ccgatatcct gctgtatgtg gtcatcccaa ccaggcgctc
     1921 cacagccgag gccttccaga tcgtgctgtc tcacctgctg ggcgacgcag gaagcccata
     1981 tctgatcggc ctgatctccg atcgcctgcg gagaaattgg cccccttctt tcctgagcga
     2041 gtttcgggcc ctgcagttca gcctgatgct gtgcgccttt gtgggcgccc tgggaggagc
     2101 agccttcctg ggcaccgcca tctttatcga ggccgacagg cgccgggcac agctgcacgt
     2161 gcagggactg ctgcacgagg caggctccac cgacgataga atcgtggtgc cccagagagg
     2221 caggtccaca agggtgcctg tggcctctgt gctgatcgac ccagctttct tgtacaaagt
     2281 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2341 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2401 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2461 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2521 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2581 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2641 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2701 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2761 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2821 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2881 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2941 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3001 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3061 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3121 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3181 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3241 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3301 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3361 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3421 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3481 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3541 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3601 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3661 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3721 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3781 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3841 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3901 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3961 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4021 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4081 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
