LOCUS       sequence-355049-        4779 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 4779)
  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 4779)
  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-SV2A_STOP
FEATURES             Location/Qualifiers
     source          1..4779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2336..2435)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2452..2471)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2453..2471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2476..2492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2476..2492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2594..2613)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2605..3414
                     /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(2672..2691)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3561..4149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4050..4069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4303..4320
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4479..4506)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4598..4684)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4733..4755
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4747..4764
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4748..4764
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatgga ggagggcttc
      121 cgggatagag ccgcctttat caggggcgcc aaggacatcg ccaaggaggt gaagaagcac
      181 gccgccaaga aggtggtgaa gggcctggat cgggtgcagg acgagtactc tcggagatcc
      241 tattctagat tcgaggagga ggacgatgac gatgactttc cagcccccag cgatggctac
      301 tatcgcggcg agggcacaca ggacgaggag gagggaggag caagctccga tgcaaccgag
      361 ggacacgacg aggatgacga gatctacgag ggagagtatc agggaatccc aagggcagag
      421 tccggaggca agggcgagag aatggccgat ggagcacctc tggccggcgt gaggggagga
      481 ctgtctgacg gcgagggccc ccctggcggc aggggcgagg cccagaggcg caaggagagg
      541 gaggagctgg cacagcagta cgaggcaatc ctgagggagt gcggacacgg ccgcttccag
      601 tggacactgt attttgtgct gggactggcc ctgatggcag atggagtgga ggtgtttgtg
      661 gtgggattcg tgctgccaag cgccgagaag gatatgtgcc tgagcgactc caacaagggc
      721 atgctgggac tcatcgtcta tcttggtatg atggtgggcg ccttcctgtg gggaggactg
      781 gcagacaggc tgggccggag acagtgcctg ctgatctctc tgagcgtgaa ttccgtgttc
      841 gccttctttt ctagctttgt gcagggctac ggcaccttcc tgttttgtag gctgctgtcc
      901 ggagtgggaa tcggaggatc tatccctatc gtgttctcct atttttctga gttcctggcc
      961 caggagaaga gaggcgagca cctgagctgg ctgtgcatgt tctggatgat cggaggcgtg
     1021 tatgcagcag caatggcatg ggcaatcatc ccacactacg gctggagctt tcagatgggc
     1081 tccgcctatc agttccactc ttggcgggtg ttcgtgctgg tgtgcgcctt tccaagcgtg
     1141 ttcgcaatcg gcgccctgac cacacagcct gagtccccac gcttctttct ggagaacggc
     1201 aagcacgatg aggcctggat ggtgctgaag caggtgcacg acaccaatat gcgggccaag
     1261 ggccaccctg agagagtgtt cagcgtgacc cacatcaaga caatccacca ggaggatgag
     1321 ctgatcgaga tccagtccga caccggcaca tggtaccagc ggtggggcgt gagagccctg
     1381 agcctgggag gacaagtgtg gggaaacttt ctgtcctgct tcggcccaga gtataggcgc
     1441 atcaccctga tgatgatggg cgtgtggttt acaatgtcct tctcttacta tggcctgacc
     1501 gtgtggttcc cagatatgat cagacacctg caggccgtgg actacgcctc taggaccaag
     1561 gtgtttccag gagagagggt ggagcacgtg accttcaact ttacactgga gaatcagatc
     1621 cacaggggcg gccagtactt taatgataag ttcatcggcc tgcgcctgaa gagcgtgtcc
     1681 tttgaggatt ccctgttcga ggagtgctat tttgaggacg tgacctcctc taacacattc
     1741 tttcggaatt gtaccttcat caacacagtg ttttacaaca cagatctgtt cgagtacaag
     1801 ttcgtgaact ctagactgat caatagcacc ttcctgcaca acaaggaggg atgtccactg
     1861 gacgtgaccg gaacaggaga gggagcctac atggtgtatt tcgtgagctt cctgggcacc
     1921 ctggccgtgc tgcctggcaa tatcgtgtct gccctgctga tggacaagat cggcaggctg
     1981 aggatgctgg caggaagctc cgtgatgagc tgcgtgtcct gtttctttct gagcttcggc
     2041 aactctgaga gcgccatgat cgccctgctg tgcctgtttg gcggcgtgtc tatcgccagc
     2101 tggaatgccc tggatgtgct gacagtggag ctgtacccct ccgacaagcg gaccacagcc
     2161 ttcggctttc tgaacgcact gtgcaagctg gcagccgtgc tgggcatctc catctttacc
     2221 tctttcgtgg gcatcacaaa ggcagcacct atcctgttcg caagcgccgc cctggccctg
     2281 ggctctagcc tggccctgaa gctgccagag accagaggcc aggtgctgca gtaagaccca
     2341 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     2401 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     2461 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     2521 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     2581 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     2641 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     2701 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     2761 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     2821 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     2881 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     2941 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     3001 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     3061 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     3121 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     3181 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     3241 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     3301 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     3361 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     3421 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     3481 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     3541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4201 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4261 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4321 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4381 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4441 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     4501 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     4561 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     4621 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     4681 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     4741 cgacgttgta aaacgacggc cagtcttaag ctcgggccc
//
