LOCUS       sequence-260434-        4707 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 4707)
  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 4707)
  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_SLC44A5
FEATURES             Location/Qualifiers
     source          1..4707
                     /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(2832..2931)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2948..2967)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2949..2967)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2972..2988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2972..2988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3090..3109)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3101..3910
                     /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(3168..3187)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4057..4645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4546..4565
                     /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 accatgaatg ataccgagaa gcctgccgac acaccatccg aggaggagga
      721 ttttggcgac cccagaacct acgatcctga cttcaagggc ccagtggcca acaggtcttg
      781 cacagatgtg ctgtgctgta tgatctttct gctgtgcatc atcggctaca tcgtgctggg
      841 actggtggca tgggtgcacg gcgatccaag gagagccgcc tatcccacag acagccaggg
      901 acacttctgc ggacagaagg gaacccctaa cgagaataag acaatcctgt tctacttcaa
      961 cctgctgcgc tgcacctctc ctagcgtgct gctgaatctg cagtgtccaa ccacacagat
     1021 ctgcgtgagc aagtgtccag agaagtttct gacctacgtg gagatgcagc tgctgtatac
     1081 aaaggataag tcctactggg aggactatcg gcagttctgt aagaccacag ccaagccagt
     1141 gaagtctctg acccagctgc tgctggacga tgactgcccc acagccatct tccccagcaa
     1201 gccttttctg cagcggtgct tccctgattt ttccaccaag aacggcaccc tgacaatcgg
     1261 ctctaagatg atgttccagg acggcaatgg aggaacacgg tccgtggtgg agctgggaat
     1321 cgcagcaaac ggcatcaata agctgctgga tgccaagtct ctgggcctga aggtgttcga
     1381 ggactacgcc agaacctggt attggatcct gatcggcctg acaatcgcca tggtgctgtc
     1441 ttggatcttc ctgatcctgc tgaggtttat cgccggctgc ctgttctggg tgtttatgat
     1501 cggcgtgatc ggcatcatcg gctacggcat ctggcactgt taccagcagt ataccaacct
     1561 gcaggagaga cccagctccg tgctgaccat ctacgatatc ggcatccaga caaatatctc
     1621 catgtatttt gagctgcagc agacctggtt caccttcatg atcatcctgt gcatcatcga
     1681 agtgatcgtg atcctgatgc tgatcttcct gaggaatcgc atccgggtgg ccatcatcct
     1741 gctgaaggag ggcagcaagg ccatcggcta cgtgccttcc accctggtgt atcccgccct
     1801 gaccttcatc ctgctgagca tctgcatctg ttactgggtg gtgaccgccg tgttcctggc
     1861 cacatccgga gtgccagtgt ataaagtgat cgcaccagga ggacactgca tccacgagaa
     1921 ccagacctgt gaccccgaga tctttaatac cacagagatc gcaaaggcat gccctggcgc
     1981 cctgtgcaac ttcgcctttt acggcggcaa gagcctgtac caccagtata tccccacctt
     2041 ccacgtgtat aacctgttcg tgtttctgtg gctgatcaat tttgtgatcg ccctgggaca
     2101 gtgcgcactg gcaggagcct tcgccaccta ctattgggcc atgaagaagc ctgatgacat
     2161 cccaaggtac cccctgttca ccgcctttgg cagagccatc aggtatcaca caggctccct
     2221 ggcctttggc tctctgatca tcgccctgat ccagatgttc aagatcgtgc tggagtacct
     2281 ggaccaccgc ctgaagcgga cccagaacac actgagcaag ttcctgcagt gctgtctgcg
     2341 ctgctgtttt tggtgcctgg agaatgccat caagttcctg aaccggaatg cctacatcat
     2401 gatcgccatc tatggcagaa acttctgtag gtccgccaag gatgccttta acctgctgat
     2461 gagaaatgtg ctgaaggtgg ccgtgaccga cgaggtgaca tacttcgtgc tgtttctggg
     2521 caagctgctg gtggcaggat ctatcggcgt gctggccttt ctgttcttta cccagaggct
     2581 gcccgtgatc gcacagggac ctgccagcct gaactactat tgggtgccac tgctgacagt
     2641 gatcttcggc tcttacctga tcgcccacgg cttctttagc gtgtatgcca tgtgcgtgga
     2701 gaccatcttc atctgttttc tggaggatct ggagaggaac gacggatcca ccgcaaggcc
     2761 atacttcgtg acacctaatc tgcacggcat cctgatcaag aagcagctgg tgccccagaa
     2821 gcagaaggag gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2881 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2941 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3001 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     3061 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     3121 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     3181 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     3241 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     3301 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     3361 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     3421 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     3481 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     3541 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     3601 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     3661 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3721 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3781 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3841 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3901 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     3961 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     4021 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4081 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4141 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4201 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4261 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4321 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4381 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4441 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4501 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     4561 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     4621 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4681 cttttgctgg ccttttgctc acatgtt
//
