LOCUS       sequence-261918-        4668 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 4668)
  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 4668)
  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_SLC44A2
FEATURES             Location/Qualifiers
     source          1..4668
                     /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(2793..2892)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2909..2928)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2910..2928)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2933..2949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3051..3070)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3062..3871
                     /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(3129..3148)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /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 accatgggcg acgagcggcc ccactactat ggcaagcacg gcacccccca
      721 gaagtacgac cccaccttca agggcccaat ctataacaga ggctgcaccg atatcatctg
      781 ctgcgtgttc ctgctgctgg caatcgtggg atacgtggca gtgggaatca tcgcatggac
      841 ccacggcgac cccagaaaag tgatctaccc tacagatagc cgcggcgagt tctgtggcca
      901 gaagggcacc aagaacgaga ataagccata cctgttctac ttcaacatcg tgaagtgcgc
      961 ctcccccctg gtgctgctgg agtttcagtg tccaaccccc cagatctgcg tggagaagtg
     1021 tcccgacagg tacctgacat atctgaatgc ccggagctcc agagatttcg agtactataa
     1081 gcagttttgc gtgcctggct tcaagaacaa taagggagtg gcagaggtgc tgcaggacgg
     1141 cgattgccct gccgtgctga tcccctctaa gcctctggcc cggagatgtt ttccagccat
     1201 ccacgcctac aagggcgtgc tgatggtggg caacgagacc acatatgagg acggccacgg
     1261 cagccggaag aacatcaccg atctggtgga gggcgccaag aaggccaatg gcgtgctgga
     1321 ggcaaggcag ctggcaatgc gcatcttcga ggactacaca gtgtcctggt attggatcat
     1381 catcggcctg gtcatcgcca tggccatgtc tctgctgttt atcatcctgc tgagattcct
     1441 ggccggcatc atggtgtggg tcatgatcat catggtcatc ctggtgctgg gctacggcat
     1501 ctttcactgc tacatggagt atagcaggct gaggggagag gcaggatctg acgtgagcct
     1561 ggtggatctg ggctttcaga ccgatttccg ggtgtacctg cacctgagac agacatggct
     1621 ggcctttatg atcatcctgt ccatcctgga agtgatcatc atcctgctgc tgatcttcct
     1681 gcggaagaga atcctgatcg ccatcgccct gatcaaggag gcatccaggg cagtgggata
     1741 cgtgatgtgc tctctgctgt atcctctggt gaccttcttt ctgctgtgcc tgtgcatcgc
     1801 ctactgggcc tctaccgccg tgtttctgag cacatccaac gaggccgtgt ataagatctt
     1861 tgacgatagc ccttgcccat tcaccgccaa gacatgtaat cctgagacat tcccatctag
     1921 caacgagagc aggcagtgcc caaatgcccg ctgtcagttc gccttttacg gaggagagtc
     1981 cggatatcac agggccctgc tgggactgca gatcttcaac gcctttatgt tcttttggct
     2041 ggccaatttt gtgctggccc tgggacaggt gaccctggca ggagccttcg ccagctacta
     2101 ttgggccctg cgcaagcctg acgatctgcc agccttcccc ctgttttctg ccttcggcag
     2161 ggccctgagg taccacacag gaagcctggc cttcggcgcc ctgatcctgg ccatcgtgca
     2221 gatcatcaga gtgatcctgg agtatctgga tcagagactg aaggccgccg agaacaagtt
     2281 tgccaagtgc ctgatgacct gtctgaagtg ctgtttctgg tgcctggaga agtttatcaa
     2341 gttcctgaac aggaatgcct acatcatgat cgccatctat ggcaccaact tctgtacatc
     2401 cgcccgcaat gccttctttc tgctgatgcg gaatatcatc agagtggccg tgctggacaa
     2461 ggtgaccgat ttcctgtttc tgctgggcaa gctgctgatc gtgggctctg tgggcatcct
     2521 ggccttcttt ttctttaccc acaggatccg catcgtgcag gacacagccc cccctctgaa
     2581 ctactattgg gtgccaatcc tgaccgtgat cgtgggcagc tacctgatcg cccacggctt
     2641 cttttccgtg tatggcatgt gcgtggatac actgttcctg tgcttcctgg aggacctgga
     2701 gcggaatgat ggctccgccg agagacccta cttcatgtcc tctaccctga agaagctgct
     2761 gaacaagaca aataagaagg ccgccgagtc tgacccagct ttcttgtaca aagttggcat
     2821 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2881 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2941 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     3001 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     3061 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     3121 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     3181 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     3241 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     3301 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     3361 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     3421 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     3481 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3541 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3601 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3661 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3721 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3781 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3841 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3901 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3961 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
