LOCUS       sequence-263267-        4479 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 4479)
  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 4479)
  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_SLC27A4
FEATURES             Location/Qualifiers
     source          1..4479
                     /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(2604..2703)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2720..2739)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2721..2739)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2744..2760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2862..2881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2873..3682
                     /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(2940..2959)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3829..4417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4318..4337
                     /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 accatgctgc tgggagcatc tctggtgggc gtgctgctgt ttagcaagct
      721 ggtgctgaag ctgccctgga cacaagtggg cttctccctg ctgtttctgt acctgggctc
      781 tggcggctgg cggttcatca gagtgtttat caagaccatc cggagagata tctttggcgg
      841 cctggtgctg ctgaaggtga aggccaaggt gaggcagtgc ctgcaggaga ggaggaccgt
      901 gccaatcctg ttcgcctcta cagtgcggag acaccccgac aagaccgccc tgatcttcga
      961 gggcaccgat acacactgga catttcgcca gctggacgag tacagctcct ctgtggcaaa
     1021 cttcctgcag gcaaggggac tggcaagcgg cgatgtggcc gccatcttca tggagaacag
     1081 aaatgagttt gtgggactgt ggctgggaat ggcaaagctg ggagtggagg ccgccctgat
     1141 caacaccaat ctgaggcgcg atgccctgct gcactgcctg accacaagca gggcacgcgc
     1201 cctggtgttt ggatctgaga tggcaagcgc catctgcgag gtgcacgcat ccctggaccc
     1261 ttctctgagc ctgttctgtt ccggatcttg ggagccagga gcagtgccac cttccaccga
     1321 gcacctggac ccactgctga aggatgcccc caagcacctg ccttcttgtc cagacaaggg
     1381 cttcacagat aagctgtttt acatctatac ctctggaacc acaggactgc caaaggcagc
     1441 aatcgtggtg cacagcaggt actatcgcat ggccgccctg gtgtactatg gcttccggat
     1501 gagacccaac gacatcgtgt acgattgcct gcctctgtat cactccgccg gcaatatcgt
     1561 gggcatcggc cagtgtctgc tgcacggcat gacagtggtc atccggaaga agttcagcgc
     1621 ctccagattt tgggacgatt gcatcaagta caactgtacc atcgtgcagt atatcggcga
     1681 gctgtgcagg tacctgctga accagccacc aagggaggca gagaatcagc accaggtgcg
     1741 gatggcactg ggaaacggac tgagacagag catctggaca aatttcagct cccggtttca
     1801 catcccccag gtggccgagt tttacggcgc caccgagtgc aactgtagcc tgggcaattt
     1861 cgactcccaa gtgggcgcct gtggcttcaa ctccagaatc ctgagcttcg tgtacccaat
     1921 ccggctggtg agagtgaatg aggataccat ggagctgatc aggggaccag acggcgtgtg
     1981 catcccatgt cagcctggag agccaggaca gctggtggga cgcatcatcc agaaggaccc
     2041 tctgcggaga tttgatggct acctgaatca gggcgccaac aataagaaga tcgccaagga
     2101 cgtgttcaag aagggcgatc aggcctatct gacaggcgac gtgctggtca tggatgagct
     2161 gggctacctg tatttcaggg accgcaccgg cgataccttc cggtggaagg gcgagaacgt
     2221 gagcaccaca gaggtggagg gcaccctgtc caggctgctg gacatggcag atgtggccgt
     2281 gtacggagtg gaggtgccag gaacagaggg aagggcagga atggcagcag tggcatcccc
     2341 taccggaaat tgtgatctgg agagatttgc ccaggtgctg gagaaggagc tgcccctgta
     2401 cgccaggcct atcttcctgc gcctgctgcc tgagctgcac aagaccggca catataagtt
     2461 ccagaagacc gagctgagga aggagggctt tgaccctgcc atcgtgaagg atccactgtt
     2521 ctatctggac gcccagaagg gcaggtacgt gccactggac caggaggcct atagccgcat
     2581 ccaggccggc gaggagaagc tggacccagc tttcttgtac aaagttggca ttataagaaa
     2641 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2701 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2761 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2821 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2881 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2941 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3001 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3061 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3121 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3181 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3241 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3301 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3361 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3421 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3481 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3541 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3601 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3661 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3721 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3781 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4441 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
