LOCUS       sequence-263873-        3846 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 3846)
  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 3846)
  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_SLC35B2
FEATURES             Location/Qualifiers
     source          1..3846
                     /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(1971..2070)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2087..2106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2088..2106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2111..2127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2111..2127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2229..2248)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2240..3049
                     /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(2307..2326)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3196..3784
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3685..3704
                     /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 accatggacg caaggtggtg ggcagtggtg gtgctggccg ccttccctag
      721 cctgggagca ggaggagaga ccccagaggc accacctgag tcctggacac agctgtggtt
      781 ctttaggttc gtggtgaacg cagcaggata cgcaagcttt atggtgcccg gctacctgct
      841 ggtgcagtat tttcggagaa agaactatct ggagaccgga aggggactgt gcttcccact
      901 ggtgaaggcc tgcgtgttcg gcaatgagcc taaggcatcc gacgaggtgc ctctggcacc
      961 aagaacagag gccgccgaga ccacacctat gtggcaggcc ctgaagctgc tgttctgcgc
     1021 caccggcctg caggtgtcct acctgacatg gggcgtgctg caggagagag tgatgacccg
     1081 cagctatgga gcaaccgcca catccccagg agagaggttc acagattctc agtttctggt
     1141 gctgatgaac cgcgtgctgg ccctgatcgt ggcaggactg tcttgcgtgc tgtgcaagca
     1201 gccaaggcac ggagcaccaa tgtaccgcta tagcttcgcc tctctgagca atgtgctgag
     1261 ctcctggtgt cagtacgagg ccctgaagtt cgtgtccttt ccaacccagg tgctggccaa
     1321 ggcctctaaa gtgatccccg tgatgctgat gggcaagctg gtgtccaggc gctcttacga
     1381 gcactgggag tatctgaccg ccacactgat ctccatcggc gtgtctatgt tcctgctgtc
     1441 tagcggacca gagccacggt cctctcctgc aaccacactg agcggcctga tcctgctggc
     1501 cggctacatc gccttcgaca gctttacctc caactggcag gatgccctgt ttgcctataa
     1561 gatgagctcc gtgcagatga tgttcggcgt gaatttcttt tcttgcctgt ttaccgtggg
     1621 aagcctgctg gagcagggcg ccctgctgga gggcacacgg ttcatgggca gacactccga
     1681 gtttgccgcc cacgccctgc tgctgtctat ctgcagcgcc tgtggccagc tgttcatctt
     1741 ttacaccatc ggccagttcg gcgccgccgt gtttaccatc atcatgacac tgcggcaggc
     1801 cttcgccatc ctgctgtctt gtctgctgta cggccacacc gtgacagtgg tgggaggact
     1861 gggagtggca gtggtgtttg ccgccctgct gctgcgggtg tatgcaaggg gcagactgaa
     1921 gcagagaggc aagaaggccg tgcccgtgga gtcccctgtg cagaaggtgg acccagcttt
     1981 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2041 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2101 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2161 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2221 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2281 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2341 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2401 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2461 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2521 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2581 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2641 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2701 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2761 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2821 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2881 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2941 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3001 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3061 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3121 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3181 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3241 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3301 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3361 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3421 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3481 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3541 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3601 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3661 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3721 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3781 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3841 catgtt
//
