LOCUS       sequence-312440-        3756 bp    DNA     circular SYN 15-APR-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 3756)
  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 3756)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC35B3_STOP
FEATURES             Location/Qualifiers
     source          1..3756
                     /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(1881..1980)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1997..2016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1998..2016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2021..2037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2021..2037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2139..2158)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2150..2959
                     /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(2217..2236)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3106..3694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3595..3614
                     /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 accatggacc tgacccagca ggccaaggat atccagaaca tcaccgtgca
      721 ggagacaaat aagaacaata gcgagtccat cgagtgctcc aagatcacca tggacctgaa
      781 gttcaacaat tctcgcaagt acatcagcat cacagtgccc tccaagaccc agacaatgtc
      841 tcctcacatc aagagcgtgg acgatgtggt ggtgctgggc atgaacctga gcaagtttaa
      901 taagctgacc cagttcttta tctgcgtggc cggcgtgttc gtgttttatc tgatctacgg
      961 ctatctgcag gagctgatct tctccgtgga gggctttaag tcatgtggat ggtaccttac
     1021 tctggtgcag ttcgcctttt attccatctt cggcctgatc gagctgcagc tgatccagga
     1081 taagcggaga aggatcccag gcaagacata catgatcatc gcctttctga ccgtgggcac
     1141 aatgggcctg agcaacacct ccctgggcta cctgaattat cctacacagg tcatcttcaa
     1201 gtgctgtaag ctgatcccag tgatgctggg cggcgtgttt atccagggca agcggtataa
     1261 cgtggccgac gtgagcgccg ccatctgcat gtccctgggc ctgatctggt tcaccctggc
     1321 cgattctacc acagccccca actttaatct gacaggcgtg gtgctgatca gcctggccct
     1381 gtgcgcagac gccgtgatcg gcaatgtgca ggagaaggcc atgaagctgc acaacgcctc
     1441 taatagcgag atggtgctgt actcctattc tatcggcttc gtgtacatcc tgctgggact
     1501 gacctgcaca agcggactgg gaccagccgt gaccttttgt gccaagaacc ccgtgagaac
     1561 atacggctat gccttcctgt tttccctgac cggctatttc ggcatctctt ttgtgctggc
     1621 cctgatcaag atcttcggcg ccctgatcgc agtgaccgtg accacaggca ggaaggccat
     1681 gacaatcgtg ctgtccttta tcttctttgc caagcccttc acatttcagt acgtgtggtc
     1741 tggcctgctg gtggtgctgg gcatcttcct gaacgtgtac agcaagaata tggacaagat
     1801 ccggctgcct tccctgtatg atctgatcaa taagtctgtg gaggcaagga agagccggac
     1861 cctggcacag acagtgtaag acccagcttt cttgtacaaa gttggcatta taagaaagca
     1921 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     1981 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2041 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa taatatcatc
     2101 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2161 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2221 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2281 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2341 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2401 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2461 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2521 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2581 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2641 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2701 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2761 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2821 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     2881 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     2941 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3001 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3061 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3121 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3181 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3721 gttcctggcc ttttgctggc cttttgctca catgtt
//
