LOCUS       sequence-260462-        5922 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 5922)
  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 5922)
  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_SLC9C2
FEATURES             Location/Qualifiers
     source          1..5922
                     /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(4047..4146)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4163..4182)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4164..4182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4187..4203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4187..4203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4305..4324)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4316..5125
                     /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(4383..4402)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5272..5860
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5761..5780
                     /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 accatgagct cctacttttg ggcccagaat gagtccaaca ggccagacct
      721 gctgtgcgga cagccagccg attatctggt ggaggagaag cacttcacca ccctggtgtg
      781 ctttatcgtg gtgctgggcg gcctgctgaa gatgtgcctg aagaattgtg aagtgatcgt
      841 gctgaccatc ctgagcctgt ccggcttcgt gatcggccac atggcctaca actccgtgga
      901 ggtgcaccag atcgtgtatc ccctgctgag aacctctagc ttcagcctgt actcctattt
      961 ttctcctctg atcatcttca tggtggccct ggacgtggag ttttacacac tgaagaagat
     1021 gttctggcag gtgctgctga caggcctgat ctctttcagc accgccagca tcatcatcgg
     1081 ctatgtggtc atcaagttca acaaggacag ctgggatctg cagtcctgcc tgctgttctc
     1141 tatcaccctg ggcatcatcg accctctgag aagcgtgaac tccctgaaga caatcggcat
     1201 cagcaagatc tacatcgatc tgatcagggg cgagtccctg atcatctgtt ctatcgccag
     1261 catcttcttt ggcaacttcc ggggcaaccg catccacttc agcatcttta gggacctgca
     1321 cgtgggcatc gagctgtctt acgatatcct gggcagcatc atcttcggct attggtgcgc
     1381 caagatcatc cagtgtatcc tggccgacgt gttttctaat atgctgacaa acatcatcct
     1441 gtgcttcagc atggtgtaca tgaccttcta tatcgtggag tttctgggaa tgtctggcac
     1501 actggccctg gcagcagtgg gcctgaatct ggatagcctg acattcaagc ccaagatcga
     1561 gctggtcatc accaagttcc tgcgcatctt ttccagcgtg tacgagcacc tgatctatgc
     1621 cttctttggc atcgtgatcg gctgtggcga gctgtctcac tacgagtttc acaccatccc
     1681 tttcatcttt atcctgttca ccacagtgaa cctggtgcgc ctgctgacaa tcctgctggt
     1741 gtccccaatc ctgatgcact ctaattacga gtataactgg agatggggag tggtcatcac
     1801 ctggagcggc atcaagggcg tgtttaatct gctgtgggcc cctgacgtgt acaacctggc
     1861 cgagcggaag gtggaggtgc cacagatgtt catcctgtac gtgcaggtca tcagcctgct
     1921 gacaatgggc atcaatagct atgtgatgac ccagtccgcc cgcaagctgg atctgtgcgt
     1981 gctgtccctg ccccggcaga tgatcctgca gaatgccacc cagcacatcc aggagatcgt
     2041 gcagaacacc atcacactgt ttaagaccga gaagatcctg acaaacgtga attggaccct
     2101 ggtggaggac aagacccgca tcgagtacat ccctttctcc cacgtgtctc acaacgacat
     2161 gaagacagag tccaccacag atgaggccct gatggaggag gcaaggctgc acgtggcagc
     2221 catccagatg agctcctttg agaagcagag aaacaatggc atcctggaga tcgaggcagc
     2281 aaggatcctg atcggagcag ccaagtgcta ctattctatc cagggcaagt tcatgagcat
     2341 ctacgacgtg agcacctata tgcggacaag aagctggctg atcaagttta agaatgtgct
     2401 gaccttcctg gagtactgta tcgagaagat ccactttatc ccccctgaga gcaacacctt
     2461 cctgacattc atctttcaca tcgtgttctc cgaggagttt gagtatacag gccagatcat
     2521 caacctgatc tacatctatc ctatgatcat ccacctgtgg ccaatggcca gaggcctgaa
     2581 cgtgagcgcc ctgatcagca tcaactacta tttcatgttt ctgtacgtgc tggagagcac
     2641 cctgaagatc atcatcctga agaggaagta tttccagcag tgctggaata cactggagtt
     2701 ctttatcctg gtcatcggca tcatcgacat cttctgcgtg tactttgtga agctgcgccc
     2761 cgataacctg gccctgatcc agctgaccgt gatcatgggc tatctgcgga tcatcagatt
     2821 cctgcccctg tttaagatca tcgtgcctat cctgatcaga atcgccgacg tgcagatcaa
     2881 gaagaggctg tccctgatgt actctatcac caagggctat atcaagtccc aggaggatgc
     2941 caagctgctg atcaagcaga tcgccgtgtg cgagtctatc taccagaagc tgtgcgagat
     3001 cctggagaca aataagcagg acgccgtgaa ggagctggtg ctgatggagc acgagggaag
     3061 ggacgtggtc atcgccctga agaccaagca ggccatccgg aatgtgatcg ccaaggccct
     3121 gaagaacctg acatttctgt gctcccgggg catcatcgac aagcacgaag tgatcgagat
     3181 caataaggtg ctgctgaaga agctgaaggc cctgaacaat ttcccaaagg ccatcccacc
     3241 ccctaccccc gatatctacc tgcacaacat catctggctg gagggcaagg acgtgctgat
     3301 cgatttcttt aaggagagag ccaagctggc ctgctttgac tccggcgata ccatctgtaa
     3361 gggcggcgag atgccacagg gcatctatct gatcatctct ggcatggcca tcctgcactc
     3421 cctgtctccc accttcggca tcgagtccaa ccagaggtgc gacaggggat ctagggatat
     3481 gttcacagag ttttgcacca caggcgacat catcggcgag ctgagctgtc tgctgaagag
     3541 ggagatcgag tacacagtga tctgcgagac ctctctgcag gcctgtttta tcagcctgga
     3601 ggacctgtac gagggcttcg atgccttttg gccatccctg gagtataaga tctggctgaa
     3661 gctggccctg tctaccgcct accagtattt cgagtctagc ctgatcgacg aggatctgcg
     3721 ctttcagaat tgcgtgatgt tcaaccaggc ctacgtggag accctgtcct cttatagcga
     3781 catgatcatc gataacatga caatgaagtt cgtgatcatc gtgtacggct ccgtgatcga
     3841 caccaagaca gaggagccat atttcgcccc ctgcatcatc cctaccacat gtgagcaggt
     3901 gcagggcacc agcgatctgt ccaagctgct gatcatccag gcctccgagc tgacacagag
     3961 aaactctaat accaacgtga tggccagcgt gaacacagtg ttcgagcagc ccggcaagaa
     4021 tatcaacggc aggcagaaga tgagcgaccc agctttcttg tacaaagttg gcattataag
     4081 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     4141 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     4201 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     4261 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     4321 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     4381 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     4441 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     4501 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     4561 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     4621 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     4681 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     4741 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     4801 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     4861 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     4921 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4981 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     5041 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     5101 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     5161 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     5221 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5281 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5341 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5401 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5461 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5521 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5581 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5641 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5701 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5761 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5821 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5881 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
