LOCUS       sequence-358240-        4218 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 4218)
  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 4218)
  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-SLC22A2_STOP
FEATURES             Location/Qualifiers
     source          1..4218
                     /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(1775..1874)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1891..1910)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1892..1910)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1915..1931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1915..1931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2033..2052)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2044..2853
                     /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(2111..2130)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3000..3588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3489..3508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3742..3759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3918..3945)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4037..4123)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4172..4194
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4186..4203
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4187..4203
                     /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 ccaccatgcc taccacagtg
      121 gacgatgtgc tggagcacgg cggcgagttt cacttctttc agaagcagat gttctttctg
      181 ctggccctgc tgagcgccac attcgcccca atctacgtgg gcatcgtgtt tctgggcttc
      241 acccctgacc acaggtgcag atccccagga gtggcagagc tgagcctgag atgtggatgg
      301 tccccagcag aggagctgaa ctacaccgtg ccaggacctg gaccagcagg agaggcatct
      361 cctagacagt gccggagata tgaggtggat tggaaccaga gcacatttga ctgcgtggat
      421 ccactggcct ctctggacac caataggagc aggctgccac tgggaccttg cagggatggc
      481 tgggtgtatg agacccccgg cagctccatc gtgacagagt tcaacctggt gtgcgccaat
      541 agctggatgc tggacctgtt ccagtctagc gtgaacgtgg gcttctttat cggctctatg
      601 agcatcggct acatcgccga tcggtttggc agaaagctgt gcctgctgac cacagtgctg
      661 atcaatgcag cagcaggcgt gctgatggca atctccccca cctatacatg gatgctgatc
      721 ttcaggctga tccagggcct ggtgtctaag gccggctggc tgatcggcta catcctgatc
      781 accgagtttg tgggcaggcg ctatcggaga acagtgggca tcttctacca ggtggcctat
      841 accgtgggac tgctggtgct ggcaggagtg gcctacgccc tgcctcactg gcgctggctg
      901 cagttcacag tgtccctgcc aaacttcttt ttcctgctgt actattggtg tatcccagag
      961 tccccccggt ggctgatctc tcagaacaag aatgccgagg ccatgagaat catcaagcac
     1021 atcgccaaga agaacggcaa gtccctgcca gcatctctgc agcggctgag actggaggag
     1081 gagacaggca agaagctgaa tccatctttt ctggacctgg tgaggactcc ccagatccgc
     1141 aagcacacaa tgatcctgat gtacaactgg ttcacctcct ctgtgctgta tcagggcctg
     1201 atcatgcaca tgggcctggc cggcgacaat atctacctgg acttcttcta ctccgccctg
     1261 gtggagtttc cagccgcctt catgatcatc ctgaccatcg acaggatcgg caggcgctac
     1321 ccatgggcag caagcaatat ggtggcagga gcagcatgcc tggcatccgt gtttatcccc
     1381 ggcgatctgc agtggctgaa gatcatcatc agctgcctgg gcaggatggg catcacaatg
     1441 gcctacgaga tcgtgtgcct ggtgaacgcc gagctgtatc ctaccttcat ccgcaatctg
     1501 ggcgtgcaca tctgcagctc catgtgcgac atcggcggca tcatcacccc ctttctggtg
     1561 tacagactga caaacatctg gctggagctg cctctgatgg tgttcggcgt gctgggactg
     1621 gtggcaggag gactggtgct gctgctgcca gagaccaagg gcaaggccct gcctgagaca
     1681 atcgaggagg ccgagaacat gcagaggcct cgcaagaata aggagaagat gatctatctg
     1741 caggtgcaga agctggatat cccactgaat taagacccag ctttcttgta caaagttggc
     1801 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     1861 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     1921 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1981 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     2041 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     2101 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2161 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2221 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2281 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2341 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2401 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2461 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2521 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2581 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2641 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2701 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2761 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2821 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     2881 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     2941 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3001 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3061 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     3781 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     3841 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     3901 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3961 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     4021 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     4081 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     4141 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     4201 agtcttaagc tcgggccc
//
