LOCUS       sequence-479330-        4164 bp    DNA     circular SYN 02-AUG-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 4164)
  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 4164)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 2, 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-SLC9B2_STOP
FEATURES             Location/Qualifiers
     source          1..4164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(690..789)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(805..822)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(805..821)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(814..836)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      885..971
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1063..1090
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1249..1266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1420..2008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1500..1519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2155..2964)
                     /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     2878..2897
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2956..2975
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3077..3093
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3077..3093
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3098..3117
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3098..3116
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3134..3233
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tcccagcagg tagtgtgcca gcagggcgct ggtacatgcc tccacgatgc aaggtcccat
       61 agacaggcgc acgcacacgc ccttcagctt cttcagggcc ttagaatcca ggcccaggcc
      121 ggctctcacc aggatgatgg acagggcgat gctccgcagg gagctagacc acttgtgctt
      181 gatctgcaca ttgtcgttga tcacggggat attcctgatc aggaagcctg ccagcagcat
      241 tcccagcagg gaaggcaggg gtggcagtgt gggcagcttg atcaggccca gcagcttgcc
      301 gccgatgatg gcacagtaaa acaggatgat gatgccgaac aggtttcctc ctggcaggca
      361 ctcagatccg gtgatggacc acaccacggc ccacagcagc acgatgattg tcacattggt
      421 gatcactctg tccagcagtc cgtgaggtgg gcatgccagc atctgtctca gccgctgcac
      481 gtggtttgcc tctgttgggg tctcctgcag cttcttctcg gagctcttca gcaggatgct
      541 gccctctgtt ggctcattgg catcgatgcc cttcagcttc atcacggtct cctcctgggc
      601 ctcctggtgc atagagggtg tatagttcat gccggtagat ggctcggagt cctcgtaggt
      661 gatccgctta tcctcgtcgc ccatggtgga gcctgctttt ttgtacaaag ttggcattat
      721 aaaaaagcat tgctcatcaa tgtgttgcaa cgaacaggtc actatcagtc aaaataaaat
      781 cattatttgg ggcccgagct taagactggc cgtcgtttta caacgtcgtg actgggaaaa
      841 catccatgct agcgttaacg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
      901 ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
      961 gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc
     1021 gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg
     1081 atggcctttt tgcgtttcta caaactcttc ctggctagcg gtacgcgtat taattgcgtt
     1141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2041 cgctcagtgg aacgacgcgt aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc
     2101 ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa
     2161 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2221 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2281 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2341 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2401 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     2461 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2521 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2581 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2641 taatacctgg aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg
     2701 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     2761 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     2821 tggcgcatcg ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc
     2881 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga
     2941 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     3001 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     3061 cacgggccag agctgccagg aaacagctat gaccatgtaa tacgactcac tataggggat
     3121 atcagctgga tggcaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
     3181 aattgataag caatgctttc ttataatgcc aactttgtac aagaaagctg ggtcttacac
     3241 ctgcacgctg gtctcgccct gcacctcctc atccttattc tggtgctcca ccttctgcag
     3301 cagtcttggt cccagcagtc cgatcagcag ggatccgata ggggctgtga tcaggatgct
     3361 cagaaaggcc acggtcagca catccatgcc atagtcctcc agctgcttct cgccgtgaga
     3421 ccgtgctgtg tccagggcca cggatccgat tgctgcctgc acggtggcct tgggcagcca
     3481 ggcaaagctg atgaagatct tctccttcag gttaaagccg gcgaagcaca ccatcaggaa
     3541 tgtggtcagg atgcggatca gcacggcgat tcccactgtt gccacgcaca gtcccacggt
     3601 ctcaggcctc aggcttgcga tagacacctc ggcgccgatc aggccaaaca gcaggggctg
     3661 gaagatatcc caggccacgg cgatgatctt ctccacctcg gccttctcgg atgtccagcc
     3721 catgccggcc agaaaggcca tgaccagggt gcacagtcct ccggatccag gaaatccgaa
     3781 gtgcacgcta gaaaacacgg ccagcacgct cagtcccagc accaggaatg tgcgcttgca
     3841 caccagctta tcctggtccc tagaggggaa gtactggata aagaagccca gcacggatcc
     3901 ggttgccact ccgatgacca cctccagcac gcccctcagc acattaaaca ctgtggagcc
     3961 ggtgctgaag gcgatgccca gacatgtgtt aaagccggtg atggccagga tatcgtcgaa
     4021 ggagcctgct gccatcagca gtgtaggcac tcccttctcc actccatatc ctcctccctg
     4081 cagcagcagc atgcttggca ccaccactgc aggagacacg gctcccagca caaagcccag
     4141 gatgaagccc cactgccatg gcag
//
