LOCUS       sequence-265829-        4110 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 4110)
  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 4110)
  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_SLC2A14
FEATURES             Location/Qualifiers
     source          1..4110
                     /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(2235..2334)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2351..2370)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2352..2370)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2375..2391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2375..2391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2493..2512)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2504..3313
                     /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(2571..2590)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3460..4048
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3949..3968
                     /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 accatggagt ttcacaatgg aggacacgtg tctggaatcg gaggcttcct
      721 ggtgagcctg acctccagga tgaagcctca cacactggcc gtgaccccag ccctgatctt
      781 cgccatcaca gtggccacca tcggctcctt ccagtttggc tacaacaccg gcgtgatcaa
      841 tgccccagag acaatcatca aggagtttat caataagaca ctgaccgaca aggcaaacgc
      901 accaccttcc gaggtgctgc tgacaaacct gtggtctctg agcgtggcca tcttttctgt
      961 gggcggcatg atcggctcct tctctgtggg cctgtttgtg aaccgcttcg gccggagaaa
     1021 ttctatgctg atcgtgaacc tgctggcagc aaccggagga tgcctgatgg gcctgtgcaa
     1081 gatcgccgag agcgtggaga tgctgatcct gggccggctg gtcatcggcc tgttttgcgg
     1141 cctgtgcaca ggcttcgtgc ccatgtatat cggagagatc agccctaccg ccctgagggg
     1201 agcctttggc acactgaatc agctgggcat cgtgatcggc atcctggtgg cccagatctt
     1261 cggcctggag ctgatcctgg gctccgagga gctgtggcca gtgctgctgg gctttaccat
     1321 cctgccagca atcctgcagt ccgccgccct gccttgctgt ccagagtctc ccaggttcct
     1381 gctgatcaat cgcaagaagg aggagaacgc aaccaggatc ctgcagcgcc tgtggggcac
     1441 acaggacgtg agccaggata tccaggagat gaaggacgag tctgcccgga tgagccagga
     1501 gaagcaggtg accgtgctgg agctgttccg ggtgagctcc tacagacagc ccatcatcat
     1561 cagcatcgtg ctgcagctgt cccagcagct gtctggcatc aatgccgtgt tttactattc
     1621 caccggcatc ttcaaggatg ccggcgtgca gcagcctatc tatgccacaa tctctgccgg
     1681 cgtggtgaac acaatcttta ccctgctgag cctgttcctg gtggagaggg caggcaggcg
     1741 caccctgcac atgatcggac tgggaggcat ggccttctgc tctacactga tgaccgtgag
     1801 cctgctgctg aagaaccact acaatggcat gagcttcgtg tgcatcggcg ccatcctggt
     1861 gtttgtggcc tgtttcgaga tcggcccagg ccccatccct tggtttatcg tggccgagct
     1921 gttctcccag ggaccaagac cagcagcaat ggcagtggcc ggctgtagca actggacctc
     1981 caattttctg gtgggcctgc tgttcccttc tgccgcctac tatctgggcg cctacgtgtt
     2041 catcatcttt acaggcttcc tgatcacctt tctggccttc acattcttta aggtgccaga
     2101 gacacggggc agaacctttg aggatatcac aagggccttc gagggacagg cacacggagc
     2161 agacagaagc ggcaaggatg gcgtgatggg catgaattcc atcgagcctg ccaaggagac
     2221 cacaaccaac gtggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2281 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2341 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2401 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2461 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2521 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2581 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2641 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2701 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2761 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2821 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2881 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2941 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3001 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3061 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3121 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3181 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3241 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3301 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3361 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3421 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3481 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3541 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3601 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3661 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3721 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3781 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3841 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3901 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3961 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4021 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4081 ggccttttgc tggccttttg ctcacatgtt
//
