LOCUS       sequence-261103-        4212 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 4212)
  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 4212)
  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_SLC22A1
FEATURES             Location/Qualifiers
     source          1..4212
                     /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(2337..2436)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2453..2472)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2454..2472)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2477..2493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2477..2493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2595..2614)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2606..3415
                     /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(2673..2692)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3562..4150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4051..4070
                     /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 accatgccca ccgtggacga tatcctggag caagtgggcg agagcggctg
      721 gttccagaag caggcctttc tgatcctgtg cctgctgtcc gccgccttcg cccctatctg
      781 cgtgggcatc gtgttcctgg gctttacccc agaccaccac tgccagagcc caggagtggc
      841 agagctgtct cagagatgtg gctggagccc agccgaggag ctgaactaca cagtgccagg
      901 actgggacct gcaggagagg cctttctggg ccagtgccgg agatatgagg tggactggaa
      961 ccagtctgcc ctgagctgcg tggatcctct ggcctccctg gccaccaata ggtctcacct
     1021 gccactggga ccatgccagg acggatgggt gtacgatacc ccaggcagct ccatcgtgac
     1081 agagttcaac ctggtgtgcg ccgactcttg gaagctggat ctgtttcaga gctgtctgaa
     1141 tgccggcttc ctgtttggca gcctgggcgt gggctatttc gccgatagat ttggcaggaa
     1201 gctgtgcctg ctgggaaccg tgctggtgaa cgccgtgtcc ggcgtgctga tggccttcag
     1261 ccccaattac atgtccatgc tgctgtttcg cctgctgcag ggcctggtgt ctaagggcaa
     1321 ttggatggcc ggctataccc tgatcacaga gttcgtggga tccggatcta ggaggacagt
     1381 ggccatcatg taccagatgg cctttaccgt gggactggtg gccctgacag gactggcata
     1441 tgcactgcca cactggaggt ggctgcagct ggccgtgtcc ctgcccacct tcctgtttct
     1501 gctgtactat tggtgcgtgc ctgagagccc aagatggctg ctgtcccaga agaggaacac
     1561 agaggccatc aagatcatgg accacatcgc ccagaagaat ggcaagctgc cccctgccga
     1621 cctgaagatg ctgagcctgg aggaggatgt gaccgagaag ctgagcccct ccttcgccga
     1681 cctgtttcgg acccctcggc tgagaaagag gacattcatc ctgatgtacc tgtggtttac
     1741 cgattccgtg ctgtatcagg gcctgatcct gcacatgggc gccacatccg gcaacctgta
     1801 cctggacttc ctgtattctg ccctggtgga gatcccaggc gcctttatcg ccctgatcac
     1861 aatcgatcgc gtgggccgga tctaccccat ggcaatgtct aacctgctgg caggagcagc
     1921 atgtctggtc atgatcttca tcagccccga cctgcactgg ctgaatatca tcatcatgtg
     1981 cgtgggccgc atgggcatca ccatcgccat ccagatgatc tgtctggtga acgccgagct
     2041 gtatcctaca tttgtgcgga atctgggcgt gatggtgtgc tctagcctgt gcgatatcgg
     2101 cggcatcatc acccctttca tcgtgtttcg cctgagggaa gtgtggcagg ccctgccact
     2161 gatcctgttc gccgtgctgg gactgctggc agcaggagtg accctgctgc tgcctgagac
     2221 aaagggcgtg gccctgccag agaccatgaa ggatgccgag aacctgggca gaaaggccaa
     2281 gcccaaggag aatacaatct acctgaaggt gcagacctcc gagccttctg gcacagaccc
     2341 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2401 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2461 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2521 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2581 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2641 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2701 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2761 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2821 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2881 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2941 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3001 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3061 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3121 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3181 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3241 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3301 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3361 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3421 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3481 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3541 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3601 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3661 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4021 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4081 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4141 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4201 tgctcacatg tt
//
