LOCUS       sequence-258945-        4410 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 4410)
  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 4410)
  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_SLC27A2
FEATURES             Location/Qualifiers
     source          1..4410
                     /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(2535..2634)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2651..2670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2652..2670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2675..2691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2675..2691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2793..2812)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2804..3613
                     /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(2871..2890)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3760..4348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4249..4268
                     /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 gcgccatcta caccgtgctg gcaggactgc tgttcctgcc
      721 actgctggtg aacctgtgct gtccctactt ctttcaggac atcggctatt ttctgaaggt
      781 ggcagcagtg ggccggagag tgcgctccta tggcaagagg cgccccgcaa ggaccatcct
      841 gagagccttc ctggagaagg cccgccagac accacacaag cccttcctgc tgtttcggga
      901 cgagaccctg acatacgccc aggtggatcg gagaagcaac caggtggcaa gagccctgca
      961 cgaccacctg ggactgaggc agggcgattg cgtggccctg ctgatgggga acgagccagc
     1021 atacgtctgg ctgtggctgg gactggtgaa gctgggatgc gcaatggcct gtctgaacta
     1081 caatatccgg gccaagagcc tgctgcactg ttttcagtgc tgtggcgcca aggtgctgct
     1141 ggtgtcccct gagctgcagg cagcagtgga ggagatcctg ccaagcctga agaaggacga
     1201 cgtgagcatc tactacgtga gcagaaccag caatacagac ggcatcgatt ctttcctgga
     1261 caaggtggat gaggtgagca ccgagcccat ccctgagtcc tggaggtctg aggtgacctt
     1321 ttccacacca gccctgtaca tctatacctc tggaaccaca ggactgccaa aggcagcaat
     1381 gatcacacac cagagaatct ggtacggcac cggcctgaca ttcgtgtctg gcctgaaggc
     1441 cgacgatgtg atctacatca ccctgccttt ttatcacagc gccgccctgc tgatcggaat
     1501 ccacggatgc atcgtggcag gagccaccct ggccctgaga acaaagttca gcgcctccca
     1561 gttttgggac gattgcagga agtacaacgt gacagtgatc cagtatatcg gcgagctgct
     1621 gaggtacctg tgcaactctc ctcagaagcc aaatgacagg gatcacaagg tgcgcctggc
     1681 cctgggaaat ggactgaggg gcgacgtgtg gagacagttc gtgaagaggt ttggcgatat
     1741 ctgtatctac gagttctatg ccgccaccga gggcaacatc ggctttatga attatgcaag
     1801 aaaagtggga gcagtgggaa gggtgaacta cctgcagaag aagatcatca catacgacct
     1861 gatcaagtat gacgtggaga aggatgagcc cgtgcgcgat gagaacggct attgcgtgcg
     1921 ggtgcctaag ggagaagtgg gactgctggt gtgcaagatc acccagctga caccattcaa
     1981 tggatacgca ggagcaaagg cacagaccga gaagaagaag ctgcgcgacg tgttcaagaa
     2041 gggcgatctg tacttcaact ccggcgacct gctgatggtg gatcacgaga atttcatcta
     2101 ctttcacgac cgcgtgggcg ataccttccg gtggaagggc gagaacgtgg ccaccacaga
     2161 ggtggccgac acagtgggcc tggtggattt tgtgcaggag gtgaacgtgt acggagtgca
     2221 cgtgccagac cacgagggac ggatcggcat ggcctccatc aagatgaagg agaatcacga
     2281 gttcgacggc aagaagctgt ttcagcacat cgccgattac ctgccctctt atgccaggcc
     2341 tcgcttcctg cgcatccagg ataccatcga gatcaccggc accttcaagc accggaagat
     2401 gacactggtg gaggagggct tcaaccctgc cgtgatcaag gacgccctgt actttctgga
     2461 cgataccgcc aagatgtatg tgccaatgac agaggatatc tacaatgcca tctccgccaa
     2521 gaccctgaag ctggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2581 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2641 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2701 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2761 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2821 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2881 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2941 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     3001 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     3061 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     3121 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     3181 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3241 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3301 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3361 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3421 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3481 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3541 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3601 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3661 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3721 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3781 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3841 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3901 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3961 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4021 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4081 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4141 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4201 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4261 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4321 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4381 ggccttttgc tggccttttg ctcacatgtt
//
