LOCUS       sequence-261066-        4218 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 4218)
  AUTHORS   Mocking TAM, Sijben HJ, Vermeulen YW, IJzerman AP, Heitman LH
  TITLE     MPP(+)-Induced Changes in Cellular Impedance as a Measure for 
            Organic Cation Transporter (SLC22A1-3) Activity and Inhibition.
  JOURNAL   Int J Mol Sci. 2022 Jan 21;23(3):1203. doi: 10.3390/ijms23031203.
  PUBMED    35163125
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"; journalName: "Int J Mol 
            Sci."; date: "2022-01-21"; volume: "23(3):1203. doi"; pages: " 
            10.3390/ijms23031203"
COMMENT     Sequence Label: pDONR221_SLC22A3
FEATURES             Location/Qualifiers
     source          1..4218
                     /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(2343..2442)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2459..2478)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2460..2478)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2483..2499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2483..2499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2601..2620)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2612..3421
                     /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(2679..2698)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3568..4156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4057..4076
                     /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 accatgccta gctttgacga ggccctgcag agggtgggag agtttggcag
      721 attccagcgg agagtgttcc tgctgctgtg cctgaccggc gtgacattcg cctttctgtt
      781 cgtgggagtg gtgttcctgg gaacccagcc agatcactac tggtgcaggg gaccatccgc
      841 cgccgccctg gcagagagat gtggctggtc tcccgaggag gagtggaaca ggacagcacc
      901 agcatccagg ggaccagagc cacctgagag gaggggcagg tgccagcggt acctgctgga
      961 ggcagcaaac gactctgcca gcgccaccag cgccctgtcc tgcgcagatc ctctggcagc
     1021 ctttccaaat aggtctgccc cactggtgcc atgtagggga ggatggagat atgcacaggc
     1081 acactctacc atcgtgagcg agttcgacct ggtgtgcgtg aacgcctgga tgctggatct
     1141 gacacaggcc atcctgaatc tgggctttct gaccggagcc ttcacactgg gatacgcagc
     1201 agaccggtat ggcagaatcg tgatctacct gctgtcttgc ctgggagtgg gagtgaccgg
     1261 agtggtggtg gcctttgccc ctaactttcc agtgttcgtg atctttcggt tcctgcaggg
     1321 cgtgttcggc aagggcacct ggatgacatg ttacgtgatc gtgaccgaga tcgtgggctc
     1381 caagcagcgg agaatcgtgg gcatcgtgat ccagatgttc tttacactgg gcatcatcat
     1441 cctgcccggc atcgcctact tcatccctaa ttggcagggc atccagctgg ccatcaccct
     1501 gcccagcttt ctgttcctgc tgtactattg ggtggtgcca gagtccccta ggtggctgat
     1561 cacaagaaag aagggcgaca aggccctgca gatcctgagg cgcatcgcca agtgcaacgg
     1621 caagtacctg agctccaatt atagcgagat caccgtgaca gacgaggagg tgtctaaccc
     1681 cagcttcctg gatctggtga ggacccctca gatgcgcaag tgtacactga tcctgatgtt
     1741 tgcctggttc acctctgccg tggtgtacca gggcctggtc atgcggctgg gcatcatcgg
     1801 cggcaacctg tatatcgatt tctttatcag cggagtggtg gagctgccag gcgccctgct
     1861 gatcctgctg accatcgaga gactgggccg gagactgcct tttgccgcaa gcaatatcgt
     1921 ggcaggagtg gcatgcctgg tgacagcctt cctgccagag ggaatcgcat ggctgaggac
     1981 cacagtggcc accctgggca gactgggcat cacaatggcc tttgagatcg tgtacctggt
     2041 gaactccgag ctgtatccca ccacactgag gaattttggc gtgtccctgt gctctggcct
     2101 gtgcgatttc ggcggcatca tcgccccttt tctgctgttc cgcctggcag ccgtgtggct
     2161 ggagctgcca ctgatcatct tcggcatcct ggcctccatc tgtggcggcc tggtcatgct
     2221 gctgcctgag accaagggca tcgccctgcc agagacagtg gacgatgtgg agaagctggg
     2281 cagcccacac tcctgcaagt gtggcaggaa taagaagacc cccgtgtccc gctctcacct
     2341 ggacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2401 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2461 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2521 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     2581 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     2641 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2701 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2761 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2821 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2881 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2941 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     3001 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     3061 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     3121 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     3181 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     3241 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     3301 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     3361 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     3421 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     3481 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4201 gccttttgct cacatgtt
//
