LOCUS       sequence-259481-        4080 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 4080)
  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 4080)
  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_SLC16A14
FEATURES             Location/Qualifiers
     source          1..4080
                     /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(2205..2304)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2321..2340)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2322..2340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2345..2361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2345..2361)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2463..2482)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2474..3283
                     /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(2541..2560)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3430..4018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3919..3938
                     /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 accatgtaca ccagccacga ggacatcggc tatgacttcg aggatggccc
      721 taaggataag aagacactga agccacaccc aaacatcgac ggaggatggg catggatgat
      781 ggtgctgagc tccttctttg tgcacatcct gatcatgggc tcccagatgg ccctgggcgt
      841 gctgaatgtg gagtggctgg aggagtttca ccagtctagg ggcctgaccg cctgggtgtc
      901 tagcctgagc atgggcatca cactgatcgt gggccccttc atcggcctgt ttatcaacac
      961 atgcggatgt aggcagaccg caatcatcgg aggactggtg aactctctgg gatgggtgct
     1021 gagcgcctac gcagcaaatg tgcactatct gttcatcacc tttggcgtgg cagcaggact
     1081 gggatccgga atggcctacc tgcctgccgt ggtcatggtg ggccggtatt tccagaagag
     1141 gagagccctg gcacagggac tgtctaccac aggaaccggc ttcggcacat ttctgatgac
     1201 cgtgctgctg aagtacctgt gcgccgagta tggctggaga aacgccatgc tgatccaggg
     1261 cgccgtgagc ctgaacctgt gcgtgtgcgg cgccctgatg aggcctctgt ctccaggcaa
     1321 gaaccccaat gaccctggcg agaaggatgt gagaggcctg cctgcccact ctacagagag
     1381 cgtgaagtcc accggccagc agggcaggac agaggagaag gatggaggac tgggaaacga
     1441 ggagaccctg tgcgacctgc aggcccagga gtgtccagat caggccggcc acaggaagaa
     1501 tatgtgcgcc ctgcgcatcc tgaagacagt gtcctggctg accatgcggg tgagaaaggg
     1561 cttcgaggac tggtactccg gctattttgg cacagcctct ctgttcacca accgcatgtt
     1621 tgtggccttc atcttttggg ccctgttcgc ctactcctct ttcgtgatcc ccttcatcca
     1681 cctgcccgag atcgtgaacc tgtataatct gagcgagcag aatgacgtgt tccctctgac
     1741 ctccatcatc gccatcgtgc acatctttgg caaagtgatc ctgggcgtga tcgccgacct
     1801 gccatgcatc agcgtgtgga acgtgttcct gctggccaac ttcaccctgg tgctgtccat
     1861 ctttatcctg cccctgatgc acacatacgc cggcctggcc gtgatctgtg ccctgatcgg
     1921 cttcagctcc ggctattttt ctctgatgcc agtggtgacc gaggatctgg tgggcatcga
     1981 gcacctggcc aacgcctacg gcatcatcat ctgcgccaat ggcatctctg ccctgctggg
     2041 accacctttc gccggatgga tctacgacat cacacagaag tacgatttca gcttttatat
     2101 ctgcggcctg ctgtatatga tcggcatcct gtttctgctg atccagccct gtatccggat
     2161 catcgagcag agcaggcgca agtacatgga cggcgcccac gtggacccag ctttcttgta
     2221 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2281 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2341 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2401 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2461 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2521 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2581 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2641 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2701 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2761 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2821 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2881 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     2941 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3001 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3061 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3121 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3181 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3241 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3301 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3361 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3541 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3781 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3841 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3901 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3961 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4021 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
