LOCUS       sequence-263265-        4086 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 4086)
  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 4086)
  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_SPNS3
FEATURES             Location/Qualifiers
     source          1..4086
                     /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(2211..2310)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2327..2346)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2328..2346)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2351..2367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2351..2367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2469..2488)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2480..3289
                     /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(2547..2566)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3436..4024
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3925..3944
                     /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 accatggcag gaggaatgtc tgccgagtgc ccagagcctg gaccaggagg
      721 actgcagggc cagagccccg gccctggccg gcagtgtcca cctccaatca cccccacaag
      781 ctggtccctg ccaccttgga gagcatacgt ggcagcagcc gtgctgtgct acatcaacct
      841 gctgaattat atgaactggt tcatcatcgc cggcgtgctg ctggacatcc aggaggtgtt
      901 tcagatctcc gataatcacg ccggcctgct gcagaccgtg ttcgtgtcct gtctgctgct
      961 gtctgccccc gtgttcggct atctgggcga caggcactcc cgcaaggcca caatgtcttt
     1021 cggcatcctg ctgtggtccg gagcaggact gagctcctct tttatctctc caaggtacag
     1081 ctggctgttc tttctgagca ggggaatcgt gggaaccgga tctgccagct attccaccat
     1141 cgccccaaca gtgctgggcg acctgttcgt gagggatcag cggacaagag tgctggccgt
     1201 gttctacatc tttatcccag tgggaagcgg actgggatat gtgctgggat ccgccgtgac
     1261 catgctgaca ggcaactggc ggtgggccct gagagtgatg ccttgcctgg aggccgtggc
     1321 cctgatcctg ctgatcctgc tggtgccaga cccacccagg ggagcagcag agacccaggg
     1381 agagggagcc gtgggcggct ttcggagctc ctggtgtgag gatgtgagat acctgggcaa
     1441 gaattggtcc ttcgtgtggt ctaccctggg cgtgacagcc atggcctttg tgacaggcgc
     1501 cctgggattc tgggcaccaa agtttctgct ggaggcaagg gtggtgcacg gactgcagcc
     1561 tccatgcttc caggagccat gtagcaaccc cgactccctg atctttggcg ccctgaccat
     1621 catgacaggc gtgatcggcg tgatcctggg agcagaggca gcacggagat ataagaaagt
     1681 gatccctgga gcagagccac tgatctgcgc atctagcctg ctggcaaccg caccatgtct
     1741 gtacctggcc ctggtgctgg caccaaccac actgctggca tcttacgtgt tcctgggact
     1801 gggagagctg ctgctgagct gcaattgggc cgtggtggcc gatatcctgc tgtccgtggt
     1861 ggtgccaagg tgtaggggaa ccgcagaggc cctgcagatc acagtgggac acatcctggg
     1921 cgacgcagga tccccctacc tgaccggcct gatctcctct gtgctgaggg caaggcgccc
     1981 tgatagctat ctgcagcggt tcagatctct gcagcagagc ttcctgtgct gtgcctttgt
     2041 gatcgccctg ggaggaggat gctttctgct gaccgccctg tacctggaga gggatgagac
     2101 aagggcatgg cagccagtga ccggaacacc tgacagcaat gacgtggatt ccaacgatct
     2161 ggagagacag ggcctgctgt ctggcgccgg cgccagcacc gaggagcctg acccagcttt
     2221 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2281 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2341 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2401 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2461 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2521 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2581 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2641 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2701 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2761 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2821 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2881 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2941 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3001 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3061 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3121 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3181 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3241 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3301 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3361 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3421 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3481 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3541 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3601 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3661 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3721 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3781 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3841 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3901 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3961 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4021 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4081 catgtt
//
