LOCUS       sequence-260210-        4215 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 4215)
  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 4215)
  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_SLC22A20P
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"
     regulatory      2228..2237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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 accatggcct tcaccgacct gctggatgca ctgggaagca tgggccggtt
      721 tcagctgaac cacacagccc tgctgctgct gccatgcgga ctgctggcat gtcacaactt
      781 cctgcagaac ttcaccgcag cagtgccacc tcaccactgc aggggaccag caaaccacac
      841 cgaggccagc acaaatgact ccggagcatg gctgagggca accatccctc tggatcagct
      901 gggagcacct gagccatgtc ggagattcac aaagccacag tgggccctgc tgtccccaaa
      961 cagctccatc cctggagcag caaccgaggg atgcaaggac ggatgggtgt acaatcggag
     1021 cgtgttccca agcaccatcg tgatggagtg ggacctggtg tgcgaggcac ggacactgag
     1081 agatctggcc cagtccgtgt acatggcagg cgtgctggtg ggagcagccg tgttcggctc
     1141 tctggcagat aggctgggat gcaagggacc actggtgtgg tcttatctgc agctggcagc
     1201 aagcggagca gcaaccgcct acttctctag cttttctgcc tattgcgtgt tccggttcct
     1261 gatgggcatg acattcagcg gcatcatcct gaatagcgtg tccctggtgg tggagtggat
     1321 gcctaccagg ggaaggacag tggcaggaat cctgctgggc tactctttta ccctgggaca
     1381 gctgatcctg gcaggagtgg cctatctgat caggccatgg aggtgcctgc agttcgcaat
     1441 cagcgccccc tttctgatct tctttctgta ctcctggtgg ctgccagagt cctctaggtg
     1501 gctgctgctg cacggcaagt cccagctggc cgtgcagaac ctgcagaagg tggccgccat
     1561 gaatggaagg aaggaggagg gagagcgcct gaccaaggaa gtgatgagct cctatatcca
     1621 gtccgagttc gccagcgtgt gcacatctaa cagcatcctg gacctgtttc ggacccccgc
     1681 catcagaaag gtgacatgct gtctgatggt catctggttc tccaattctg tggcctacta
     1741 tggcctggcc atggatctgc agaagttcgg cctgagcctg tacctggtgc aggccctgtt
     1801 tggcatcatc aacacccctg caatgctggt ggcaaccgca acaatgatct atgtgggcag
     1861 gcgcgcaaca gtggcatcct tcctgatcct ggccggcctg atggtcatcg ccaatatgtt
     1921 tgtgccagag ggcacccaga tcctgtgcac agcacaggcc gccctgggca agggatgcct
     1981 ggcctctagc ttcatctgcg tgtacctgtt caccggcgag ctgtatccca cagagatccg
     2041 gcagatggga atgggattcg caagcgtgca cgcaagactg ggaggactga ccgcacctct
     2101 ggtgaccaca ctgggcgagt actccaccat cctgccaccc gtgtcttttg gcgcaacagc
     2161 aatcctggca ggactggccg tgtgcgtgct gaccgagaca aggaacatgc ccctggtgga
     2221 gaccatcgcc gccatggagc ggagagtgaa ggagggctcc tctaagaagc acgtggagga
     2281 gaagagcgag gagatctccc tgcagcagct gagggcaagc cctctgaagg agacaatcga
     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
//
