LOCUS       sequence-259077-        3873 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 3873)
  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 3873)
  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
FEATURES             Location/Qualifiers
     source          1..3873
                     /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(1998..2097)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2114..2133)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2115..2133)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2138..2154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2138..2154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2256..2275)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2267..3076
                     /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(2334..2353)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3223..3811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3712..3731
                     /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 cgcgccgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     2041 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2101 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2161 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2221 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2281 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2341 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2401 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2461 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2521 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2581 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2641 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2701 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2761 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2821 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2881 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2941 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3001 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3061 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     3121 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     3181 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3241 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3301 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3361 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3421 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3481 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3541 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3601 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3661 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3721 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3781 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3841 cctggccttt tgctggcctt ttgctcacat gtt
//
