LOCUS       sequence-312032-        3876 bp    DNA     circular SYN 15-DEC-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 3876)
  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 3876)
  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-RHBG_STOP
FEATURES             Location/Qualifiers
     source          1..3876
                     /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(2001..2100)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2117..2136)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2118..2136)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2141..2157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2141..2157)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2259..2278)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2270..3079
                     /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(2337..2356)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3226..3814
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3715..3734
                     /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 gaagcccttc cagggcagca ggccggagac tgcagctgcc
      721 actgctgtgc ctgttcctgc agggagcaac cgccgtgctg ttcgccgtgt ttgtgaggta
      781 caatcacaag acagacgccg ccctgtggca ccgctctaac cacagcaatg ccgacaacga
      841 gttctacttt cggtatccca gctttcagga tgtgcacgcc atggtgttcg tgggcttcgg
      901 ctttctgatg gtgtttctgc agagatatgg cttcagctcc gtgggcttca cctttctgct
      961 ggcagccttc gccctgcagt ggtccacact ggtgcagggc ttcctgcact cttttcacgg
     1021 aggacacatc cacgtgggag tggagtccat gatcaatgcc gatttttgtg caggagccgt
     1081 gctgatctct ttcggcgccg tgctgggcaa gaccggacct acacagctgc tgctgatggc
     1141 cctgctggag gtggtgctgt ttggcatcaa cgagttcgtg ctgctgcacc tgctgggcgt
     1201 gcgggacgca ggaggctcca tgaccatcca cacattcggc gcctactttg gcctggtgct
     1261 gagcagggtg ctgtatcgcc cacagctgga gaagtccaag cacagacagg gcagcgtgta
     1321 ccactccgat ctgtttgcca tgatcggcac catcttcctg tggatctttt ggccctcttt
     1381 caatgccgcc ctgacagccc tgggagcagg acagcacagg accgccctga acacatacta
     1441 ttctctggca gcaagcaccc tgggcacatt cgccctgtcc gccctggtgg gagaggacgg
     1501 ccgcctggat atggtgcaca tccagaatgc cgccctggca ggaggagtgg tggtgggcac
     1561 ctctagcgag atgatgctga caccctttgg cgccctggca gcaggcttcc tggcaggaac
     1621 cgtgtctaca ctgggctaca agttctttac ccccatcctg gagagcaagt ttaaggtgca
     1681 ggacacatgc ggcgtgcaca acctgcacgg catgcctggc gtgctgggcg ccctgctggg
     1741 cgtgctggtg gcaggcctgg ccacccacga ggcctatggc gatggcctgg agagcgtgtt
     1801 ccctctgatc gcagagggac agaggtccgc cacctctcag gccatgcacc agctgttcgg
     1861 cctgtttgtg acactgatgt ttgcaagcgt gggaggagga ctgggaggac tgctgctgaa
     1921 gctgccattc ctggactccc cacctaggct gccagccctg agaggaccct cctctctggc
     1981 cggcgcctgg cgcgcctaag acccagcttt cttgtacaaa gttggcatta taagaaagca
     2041 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2101 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     2161 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa taatatcatc
     2221 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2281 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     2341 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     2401 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     2461 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     2521 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     2581 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     2641 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     2701 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     2761 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     2821 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     2881 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     2941 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3001 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3061 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3121 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     3181 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3241 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3301 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3361 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3421 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3481 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3541 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3601 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3661 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3721 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3781 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3841 gttcctggcc ttttgctggc cttttgctca catgtt
//
