LOCUS       sequence-374402-        4200 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 4200)
  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 4200)
  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-SLC33A1_STOP
FEATURES             Location/Qualifiers
     source          1..4200
                     /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(2325..2424)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2441..2460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2442..2460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2465..2481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2465..2481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2583..2602)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2594..3403
                     /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(2661..2680)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3550..4138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4039..4058
                     /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 accatgagcc ctaccatctc ccacaaggac agctccaggc agcggagacc
      721 aggcaatttc tcccactctc tggatatgaa gtccggacca ctgccacctg gaggatggga
      781 cgattctcac ctggacagcg ccggaaggga gggcgataga gaggccctgc tgggcgacac
      841 aggaaccggc gatttcctga aggcccctca gtcctttcgc gccgagctgt ctagcatcct
      901 gctgctgctg tttctgtatg tgctgcaggg aatcccactg ggactggcag gatctatccc
      961 actgatcctg cagtccaaga acgtgtctta cacagaccag gccttctttt ccttcgtgtt
     1021 ttggcctttc tctctgaagc tgctgtgggc cccactggtg gatgccgtgt acgtgaagaa
     1081 ttttggcagg cgcaagtcct ggctggtgcc cacacagtat atcctgggcc tgttcatgat
     1141 ctacctgtct acccaggtgg accggctgct gggcaacaca gacgatagaa cccctgatgt
     1201 gatcgccctg accgtggcct tctttctgtt cgagtttctg gccgccacac aggacatcgc
     1261 cgtggatggc tgggccctga ccatgctgtc tagggagaat gtgggctatg ccagcacatg
     1321 caactccgtg ggacagaccg caggatactt cctgggcaat gtgctgtttc tggccctgga
     1381 gagcgccgac ttctgtaaca agtatctgag gtttcagcca cagccccgcg gcatcgtgac
     1441 actgtccgat ttcctgttct tttggggcac cgtgtttctg atcaccacaa ccctggtggc
     1501 cctgctgaag aaggagaatg aggtgagcgt ggtgaaggag gagacacagg gcatcacaga
     1561 cacctacaag ctgctgttcg ccatcatcaa gatgcccgcc gtgctgacat tttgcctgct
     1621 gatcctgacc gccaagatcg gcttcagcgc cgcagatgca gtgaccggcc tgaagctggt
     1681 ggaggaggga gtgcctaagg agcacctggc cctgctggca gtgccaatgg tgcctctgca
     1741 gatcatcctg ccactgatca tctccaagta tacagccggc cctcagccac tgaatacctt
     1801 ttataaggcc atgccttaca ggctgctgct gggactggag tacgcactgc tggtgtggtg
     1861 gacacccaag gtggagcacc agggcggctt tcctatctac tattacatcg tggtgctgct
     1921 gtcttatgcc ctgcaccagg tgaccgtgta ttctatgtac gtgagcatca tggccttcaa
     1981 cgccaaggtg agcgacccac tgatcggcgg cacatacatg accctgctga acaccgtgag
     2041 caatctgggc ggcaactggc catccacagt ggccctgtgg ctggtggatc cactgaccgt
     2101 gaaggagtgc gtgggagcaa gcaaccagaa ttgtcggaca ccagacgccg tggagctgtg
     2161 caagaagctg ggaggatctt gcgtgaccgc cctggatggc tattacgtgg agagcatcat
     2221 ctgcgtgttc atcggctttg gctggtggtt ctttctgggc cccaagttca agaagctgca
     2281 ggacgagggc tcctctagct ggaagtgtaa gagaaacaat taagacccag ctttcttgta
     2341 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2401 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2461 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2521 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2581 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2641 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2701 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2761 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2821 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2881 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2941 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3001 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3061 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3121 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3181 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3241 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3301 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3361 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3421 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3481 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3541 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3601 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3661 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3721 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3781 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3841 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3901 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3961 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4021 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4081 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4141 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
