LOCUS       sequence-260113-        4215 bp    DNA     circular SYN 14-APR-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 4215)
  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 4215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SLC22A2
FEATURES             Location/Qualifiers
     source          1..4215
                     /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(2340..2439)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2456..2475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2457..2475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2480..2496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2598..2617)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2609..3418
                     /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(2676..2695)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3565..4153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4054..4073
                     /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 ccacagtgga cgatgtgctg gagcacggcg gcgagtttca
      721 cttctttcag aagcagatgt tctttctgct ggccctgctg agcgccacat tcgccccaat
      781 ctacgtgggc atcgtgtttc tgggcttcac ccctgaccac aggtgcagat ccccaggagt
      841 ggcagagctg agcctgagat gtggatggtc cccagcagag gagctgaact acaccgtgcc
      901 aggacctgga ccagcaggag aggcatctcc tagacagtgc cggagatatg aggtggattg
      961 gaaccagagc acatttgact gcgtggatcc actggcctct ctggacacca ataggagcag
     1021 gctgccactg ggaccttgca gggatggctg ggtgtatgag acccccggca gctccatcgt
     1081 gacagagttc aacctggtgt gcgccaatag ctggatgctg gacctgttcc agtctagcgt
     1141 gaacgtgggc ttctttatcg gctctatgag catcggctac atcgccgatc ggtttggcag
     1201 aaagctgtgc ctgctgacca cagtgctgat caatgcagca gcaggcgtgc tgatggcaat
     1261 ctcccccacc tatacatgga tgctgatctt caggctgatc cagggcctgg tgtctaaggc
     1321 cggctggctg atcggctaca tcctgatcac cgagtttgtg ggcaggcgct atcggagaac
     1381 agtgggcatc ttctaccagg tggcctatac cgtgggactg ctggtgctgg caggagtggc
     1441 ctacgccctg cctcactggc gctggctgca gttcacagtg tccctgccaa acttcttttt
     1501 cctgctgtac tattggtgta tcccagagtc cccccggtgg ctgatctctc agaacaagaa
     1561 tgccgaggcc atgagaatca tcaagcacat cgccaagaag aacggcaagt ccctgccagc
     1621 atctctgcag cggctgagac tggaggagga gacaggcaag aagctgaatc catcttttct
     1681 ggacctggtg aggactcccc agatccgcaa gcacacaatg atcctgatgt acaactggtt
     1741 cacctcctct gtgctgtatc agggcctgat catgcacatg ggcctggccg gcgacaatat
     1801 ctacctggac ttcttctact ccgccctggt ggagtttcca gccgccttca tgatcatcct
     1861 gaccatcgac aggatcggca ggcgctaccc atgggcagca agcaatatgg tggcaggagc
     1921 agcatgcctg gcatccgtgt ttatccccgg cgatctgcag tggctgaaga tcatcatcag
     1981 ctgcctgggc aggatgggca tcacaatggc ctacgagatc gtgtgcctgg tgaacgccga
     2041 gctgtatcct accttcatcc gcaatctggg cgtgcacatc tgcagctcca tgtgcgacat
     2101 cggcggcatc atcaccccct ttctggtgta cagactgaca aacatctggc tggagctgcc
     2161 tctgatggtg ttcggcgtgc tgggactggt ggcaggagga ctggtgctgc tgctgccaga
     2221 gaccaagggc aaggccctgc ctgagacaat cgaggaggcc gagaacatgc agaggcctcg
     2281 caagaataag gagaagatga tctatctgca ggtgcagaag ctggatatcc cactgaatga
     2341 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     2401 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     2461 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     2521 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     2581 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2641 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2701 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2761 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2821 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2881 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2941 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     3001 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     3061 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     3121 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     3181 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     3241 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     3301 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     3361 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     3421 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     3481 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     4201 ttttgctcac atgtt
//
