LOCUS       sequence-260399-        4125 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 4125)
  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 4125)
  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_SLC18A1
FEATURES             Location/Qualifiers
     source          1..4125
                     /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"
     RBS             835..843
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     protein_bind    complement(2250..2349)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2366..2385)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2367..2385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2390..2406)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2390..2406)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2508..2527)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2519..3328
                     /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(2586..2605)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3475..4063
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3964..3983
                     /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 accatgctga ggaccatcct ggacgcacca cagagactgc tgaaggaggg
      721 aagggcaagc aggcagctgg tgctggtggt ggtgttcgtg gccctgctgc tggacaatat
      781 gctgtttacc gtggtggtgc caatcgtgcc cacattcctg tacgatatgg agtttaagga
      841 ggtgaacagc tccctgcacc tgggacacgc aggatctagc ccacacgcac tggcatctcc
      901 tgccttcagc accatctttt ccttctttaa caataacaca gtggcagtgg aggagagcgt
      961 gccatccgga atcgcctgga tgaatgatac cgcatccaca atcccacctc cagcaaccga
     1021 ggcaatctct gcccacaaga ataactgcct gcagggaacc ggcttcctgg aggaggagat
     1081 cacacgggtg ggcgtgctgt ttgccagcaa ggccgtgatg cagctgctgg tgaatccctt
     1141 cgtgggccct ctgacaaaca gaatcggcta ccacatcccc atgttcgccg gctttgtgat
     1201 catgtttctg tccaccgtga tgttcgcctt ttctggcacc tatacactgc tgttcgtggc
     1261 acggacactg cagggaatcg gatcctcttt tagctccgtg gcaggactgg gaatgctggc
     1321 atccgtgtac accgacgatc acgagcgggg cagagccatg ggaacagccc tgggaggact
     1381 ggccctgggc ctgctggtgg gcgccccttt cggcagcgtg atgtatgagt ttgtgggcaa
     1441 gtccgcccca ttcctgatcc tggcctttct ggccctgctg gacggcgccc tgcagctgtg
     1501 catcctgcag ccatctaagg tgagcccaga gtccgccaag ggaaccccac tgttcatgct
     1561 gctgaaggat ccctacatcc tggtggccgc cggctctatc tgctttgcca atatgggagt
     1621 ggcaatcctg gagcctaccc tgccaatctg gatgatgcag acaatgtgca gccctaagtg
     1681 gcagctggga ctggccttcc tgccagcatc cgtgtcttat ctgatcggca ccaatctgtt
     1741 tggcgtgctg gccaacaaga tgggcaggtg gctgtgctct ctgatcggca tgctggtggt
     1801 gggcacaagc ctgctgtgcg tgccactggc ccacaatatc ttcggactga tcggacctaa
     1861 cgcaggactg ggactggcaa tcggaatggt ggactctagc atgatgccta tcatgggcca
     1921 cctggtggat ctgcgccaca ccagcgtgta cggcagcgtg tatgccatcg ccgacgtggc
     1981 cttctgcatg ggctttgcaa tcggaccatc caccggagga gcaatcgtga aggcaatcgg
     2041 cttcccctgg ctgatggtca tcacaggcgt gatcaacatc gtgtacgccc ccctgtgcta
     2101 ctatctgagg agcccccctg ccaaggagga gaagctggcc atcctgtccc aggactgtcc
     2161 tatggagaca cgcatgtatg ccacccagaa gcctacaaag gagtttccac tgggcgagga
     2221 ctctgatgag gagcccgatc acgaggagga cccagctttc ttgtacaaag ttggcattat
     2281 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     2341 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     2401 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     2461 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2521 gagccatatt caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt
     2581 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt
     2641 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2701 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     2761 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     2821 aaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2881 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     2941 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3001 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3061 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3121 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3181 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3241 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     3301 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     3361 gagcattacg ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag
     3421 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3481 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3541 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3601 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3661 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3721 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3781 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3841 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3901 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3961 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4021 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4081 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
