LOCUS       sequence-346619-        3675 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 3675)
  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 3675)
  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-SLC35F2_STOP
FEATURES             Location/Qualifiers
     source          1..3675
                     /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(1232..1331)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1348..1367)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1349..1367)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1372..1388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1372..1388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1490..1509)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1501..2310
                     /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(1568..1587)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2457..3045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2946..2965
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3199..3216
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3375..3402)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3494..3580)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3629..3651
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3643..3660
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3644..3660
                     /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 ggccgatagc
      121 cctgcaggac caggagcacc agagccactg gccgagggcg ccgccgccga gtttagctcc
      181 ctgctgcgga gaatcaaggg caagctgttc acatggaaca tcctgaagac catcgccctg
      241 ggccagatgc tgtctctgtg catctgtggc acagccatca ccagccagta cctggccgag
      301 cggtataagg tgaacacacc aatgctgcag tccttcatca attactgcct gctgtttctg
      361 atctataccg tgatgctggc ctttcggtcc ggctctgata acctgctggt catcctgaag
      421 agaaagtggt ggaagtacat cctgctggga ctggcagacg tggaggccaa ttatgtgatc
      481 gtgcgggcct accagtatac cacactgaca agcgtgcagc tgctggattg cttcggcatc
      541 cccgtgctga tggccctgtc ctggtttatc ctgcacgcca ggtaccgcgt gatccacttc
      601 atcgcagtgg ccgtgtgcct gctgggagtg ggaaccatgg tgggagcaga catcctggca
      661 ggaagggagg acaacagcgg atccgatgtg ctgatcggcg acatcctggt gctgctggga
      721 gcatccctgt acgccatctc taacgtgtgc gaggagtata tcgtgaagaa gctgagcagg
      781 caggagttcc tgggaatggt gggactgttt ggcaccatca tctccggcat ccagctgctg
      841 atcgtggagt acaaggatat cgcctctatc cactgggact ggaagatcgc cctgctgttc
      901 gtggcctttg ccctgtgcat gttctgtctg tactccttta tgcctctggt catcaaggtg
      961 acatctgcca ccagcgtgaa tctgggcatc ctgacagccg atctgtattc tctgttcgtg
     1021 ggcctgttcc tgtttggcta caagtttagc ggcctgtata tcctgtcctt caccgtgatc
     1081 atggtgggct ttatcctgta ttgttctacc cccacaagaa ccgcagagcc tgcagagtct
     1141 agcgtgccac ctgtgacaag catcggcatc gacaacctgg gcctgaagct ggaggagaat
     1201 ctgcaggaga cccactccgc cgtgctgtaa gacccagctt tcttgtacaa agttggcatt
     1261 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1321 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     1381 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1441 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     1501 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     1561 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     1621 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     1681 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     1741 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     1801 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     1861 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     1921 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     1981 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2041 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2101 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2161 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2221 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2281 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     2341 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     2401 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     2821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3061 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3121 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3181 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3241 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3301 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3361 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3421 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     3481 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3541 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     3601 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     3661 cttaagctcg ggccc
//
