LOCUS       sequence-262920-        4197 bp    DNA     circular SYN 14-APR-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 4197)
  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 4197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SPNS2
FEATURES             Location/Qualifiers
     source          1..4197
                     /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(2322..2421)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2438..2457)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2439..2457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2462..2478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2462..2478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2580..2599)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2591..3400
                     /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(2658..2677)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3547..4135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4036..4055
                     /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 accatgatgt gcctggagtg tgcatccgcc gcagcaggag gagcagagga
      721 ggaggaggcc gacgccgagc ggagaaggcg ccggagaggc gcccagaggg gcgccggcgg
      781 cagcggctgc tgtggagcaa ggggagcagg aggagcaggc gtgtccgccg ccggcgatga
      841 ggtgcagacc ctgagcggct ccgtgaggcg cgccccaaca ggcccccctg gcaccccagg
      901 cacacccggc tgtgcagcaa ccgcaaaggg accaggagca cagcagccca agcctgcctc
      961 tctgggaagg ggaaggggag cagcagcagc aatcctgagc ctgggcaacg tgctgaatta
     1021 cctggaccgg tatacagtgg ccggcgtgct gctggacatc cagcagcact tcggcgtgaa
     1081 ggatagaggc gccggcctgc tgcagtccgt gttcatctgt tcttttatgg tggccgcccc
     1141 aatcttcggc tacctgggcg atcggtttaa cagaaaagtg atcctgagct gcggcatctt
     1201 cttttggtcc gccgtgacct tcagctcctc ttttatcccc cagcagtact tctggctgct
     1261 ggtgctgtcc aggggactgg tgggaatcgg agaggcctct tatagcacca tcgcccccac
     1321 aatcatcggc gacctgttta ccaagaatac cagaacactg atgctgagcg tgttctactt
     1381 tgccatccct ctgggctccg gcctgggcta tatcacagga agctccgtga agcaggcagc
     1441 aggcgattgg cactgggcac tgagggtgag ccctgtgctg ggcatgatca ccggcacact
     1501 gatcctgatc ctggtgccag ccaccaagag gggacacgca gaccagctgg gcgatcagct
     1561 gaaggcccgg acatcctggc tgagagacat gaaggccctg atccggaaca gatcttacgt
     1621 gttctctagc ctggccacct ccgccgtgtc ttttgccaca ggcgccctgg gcatgtggat
     1681 ccctctgtat ctgcacaggg cccaggtggt gcagaagacc gccgagacat gcaattctcc
     1741 accctgtggc gccaaggaca gcctgatctt cggcgccatc acctgtttca caggctttct
     1801 gggcgtggtg accggagcag gagcaacaag gtggtgccgc ctgaagaccc agagggcaga
     1861 tccactggtg tgcgcagtgg gaatgctggg atccgccatc ttcatctgcc tgatctttgt
     1921 ggccgccaag tcctctatcg tgggcgccta catctgcatc ttcgtgggcg agaccctgct
     1981 gttttctaac tgggccatca cagccgacat cctgatgtat gtggtcatcc ctaccaggag
     2041 agcaacagca gtggccctgc agagcttcac ctcccacctg ctgggcgatg caggatcccc
     2101 atacctgatc ggctttatct ctgacctgat caggcagtct acaaaggata gccccctgtg
     2161 ggagttcctg agcctgggct atgccctgat gctgtgccct tttgtggtgg tgctgggcgg
     2221 catgttcttt ctggccaccg ccctgttctt cgtgagcgat agggcaaggg cagagcagca
     2281 agtgaatcag ctggccatgc ctccagccag cgtgaaggtg gacccagctt tcttgtacaa
     2341 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     2401 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     2461 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     2521 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     2581 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     2641 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     2701 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     2761 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     2821 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     2881 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     2941 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3001 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3061 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3121 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3181 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3241 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3301 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     3361 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     3421 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     3481 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3661 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3901 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
