LOCUS       sequence-260444-        4332 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 4332)
  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 4332)
  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_SLC22A14
FEATURES             Location/Qualifiers
     source          1..4332
                     /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(2457..2556)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2573..2592)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2574..2592)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2597..2613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2597..2613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2715..2734)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2726..3535
                     /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(2793..2812)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3682..4270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4171..4190
                     /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 accatggccg gcgaggagaa cttcaaggag gagctgcgga gccaggacgc
      721 atccagaaac ctgaatcagc acgaggtggc cggccaccct cactcttgga gcctggagat
      781 gctgctgcgg agactgaggg cagtgcacac caagcaggac gataagtttg ccaacctgct
      841 ggacgcagtg ggagagttcg gcacctttca gcagcggctg gtggccctga cattcatccc
      901 atctatcatg agcgccttct ttatgtttgc cgatcacttc gtgtttaccg cccagaagcc
      961 ttactgcaat acatcctgga tcctggccgt gggcccacac ctgtctaagg ccgagcagct
     1021 gaacctgacc atcccacagg cccccaatgg ctctttcctg acatgtttta tgtacctgcc
     1081 cgtgccttgg aacctggaca gcatcatcca gttcggcctg aatgacaccg atacatgcca
     1141 ggacggctgg atctatcccg atgccaagaa gagatctctg atcaacgagt ttgacctggt
     1201 gtgcggcatg gagaccaaga aggatacagc ccagatcatg ttcatggccg gcctgccaat
     1261 cggcagcctg atctttaggc tgatcaccga taagatgggc cgctatccag caatcctgct
     1321 gtccctgctg ggactgatca tcttcggctt tggcacagcc ttcatgaatt cttttcacct
     1381 gtacctgttc tttagattcg gcatctccca gtctgtggtg ggctatgcca tcagctccat
     1441 ctccctggca accgagtggc tggtgggaga gcacagggca cacgccatca tcctgggcca
     1501 ctgcttcttt gcagtgggag ccgtgctgct gacaggaatc gcctacagcc tgccccactg
     1561 gcagctgctg ttcctggtgg gcggcatcct ggtcatccct tttatctcct atatctggat
     1621 cctgcctgag tctccacgct ggctgatgat gaagggcaag gtgaaggagg ccaagcaggt
     1681 gctgtgctac gccgccagcg tgaacaagaa gaccatccca tccaatctgc tggacgagct
     1741 gcagctgccc cggaagaagg tgacaagagc ctccgtgctg gatttctgca agaatcggca
     1801 gctgtgcaag gtgaccctgg tcatgtcctg cgtgtggttc acagtgtctt acacctactt
     1861 caccctgagc ctgaggatga gggagctggg cgtgtctgtg cacttccggc acgtggtgcc
     1921 tagcatcatg gaggtgccag ccagactgtg ctgtatcttt ctgctgcagc agatcggcag
     1981 gaagtggagc ctggcagtga ccctgctgca ggccatcatc tggtgcctgc tgctgctgtt
     2041 cctgccagag ggagaggatg gactgaggct gaagtggcca aggtgtcctg caaccgagct
     2101 gaagagcatg acaatcctgg tgctgatgct gagagagttt tccctggccg ccaccgtgac
     2161 agtgttcttt ctgtataccg cagagctgct gcctaccgtg ctgagggcaa caggactggg
     2221 actggtgtcc ctggcatctg tggcaggcgc catcctgagc ctgaccatca tctctcagac
     2281 accaagcctg ctgcccatct tcctgtgctg cgtgctggcc atcgtggcct tttccctgtc
     2341 tagcctgctg ccagagacca gggaccagcc tctgagcgag tccctgaacc actcctctca
     2401 gatccggaat aaggtgaagg atatgaagac caaggagaca agctccgacg atgtggaccc
     2461 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2521 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2581 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2641 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2701 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2761 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2821 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2881 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2941 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3001 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     3061 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3121 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3181 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3241 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3301 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3361 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3421 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3481 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3541 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3601 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3661 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3721 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3781 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3841 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3901 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3961 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4021 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4081 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4141 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4201 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4261 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4321 tgctcacatg tt
//
