LOCUS       sequence-451007-        4215 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 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-SLC22A1_STOP
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 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 gcacataaga
     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
//
