LOCUS       sequence-339576-        3453 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 3453)
  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 3453)
  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-SLC25A35_STOP
FEATURES             Location/Qualifiers
     source          1..3453
                     /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(1010..1109)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1126..1145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1127..1145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1150..1166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1150..1166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1268..1287)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1279..2088
                     /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(1346..1365)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2235..2823
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2724..2743
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2977..2994
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3153..3180)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3272..3358)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3407..3429
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3421..3438
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3422..3438
                     /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 cttcctgatg
      121 tctggactgg cagcatgcgg agcatgcgtg ttcaccaacc ctctggaggt ggtgaagacc
      181 aggatgcagc tgcagggaga gctgcaagca ccgggtacct accaacgaca ctatagaaac
      241 gtgttccacg cctttatcac catcggcaag gtggatggac tggccgccct gcagaaggga
      301 ctggcacctg ccctgctgta ccagttcctg atgaacggaa tcaggctggg aacatacgga
      361 ctggcagagg caggaggata tctgcacacc gcagagggaa cacactcccc agcaagatct
      421 gccgcagcag gagccatggc cggcgtgatg ggcgcctacc tgggctcccc tatctatatg
      481 gtgaagacac acctgcaggc acaggcagca tctgagatcg cagtgggcca ccagtataag
      541 caccagggca tgtttcaggc cctgaccgag atcggacaga agcacggact ggtgggactg
      601 tggaggggcg ccctgggagg actgccacgc gtgatcgtgg gcagctccac ccagctgtgc
      661 acattctcta gcaccaagga cctgctgagc cagtgggaga tctttccccc tcagtcctgg
      721 aagctggccc tggtggcagc aatgatgagc ggaatcgcag tggtgctggc aatggcacca
      781 ttcgacgtgg cctgtacacg gctgtacaat cagccaaccg atgcacaggg caagggactg
      841 atgtatcggg gcatcctgga tgccctgctg cagaccgcca gaacagaggg catcttcggc
      901 atgtacaagg gcatcggagc atcctacttc cggctgggac cacacaccat cctgtctctg
      961 ttcttttggg accagctgcg cagcctgtac tataccgata caaagtaaga cccagctttc
     1021 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1081 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     1141 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1201 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1261 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
     1321 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     1381 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     1441 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     1501 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     1561 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
     1621 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     1681 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     1741 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     1801 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
     1861 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1921 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1981 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     2041 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     2101 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     2161 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2221 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2281 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2341 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2401 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2461 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2521 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2581 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2641 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2701 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2761 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2821 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2881 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2941 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3001 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3061 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3121 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3181 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3241 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3301 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3361 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3421 tgtaaaacga cggccagtct taagctcggg ccc
//
