LOCUS       sequence-259330-        4092 bp    DNA     circular SYN 14-APR-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 4092)
  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 4092)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_MFSD4A
FEATURES             Location/Qualifiers
     source          1..4092
                     /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"
     regulatory      1799..1808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2217..2316)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2333..2352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2334..2352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2357..2373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2357..2373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2475..2494)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2486..3295
                     /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(2553..2572)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3442..4030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3931..3950
                     /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 accatgggat gcgacggacg ggtgagcgga ctgctgcgga gaaacctgca
      721 gccaaccctg acatactgga gcgtgttctt ttccttcggc ctgtgcatcg cctttctggg
      781 ccctaccctg ctggatctga gatgtcagac acacagctcc ctgccacaga tctcttgggt
      841 gttctttagc cagcagctgt gcctgctgct gggatccgcc ctgggaggcg tgttcaagcg
      901 gaccctggca cagagcctgt gggcactgtt tacatctagc ctggccatct ccctggtgtt
      961 cgccgtgatc cccttttgca gagacgtgaa ggtgctggca tctgtgatgg cactggcagg
     1021 actggcaatg ggatgtatcg acaccgtggc caacatgcag ctggtgagga tgtatcagaa
     1081 ggatagcgcc gtgttcctgc aggtgctgca cttctttgtg ggctttggcg ccctgctgag
     1141 ccctctgatc gccgatccat tcctgtccga ggccaattgc ctgcccgcca acagcaccgc
     1201 caataccaca tccaggggcc acctgtttca cgtgtcccgc gtgctgggac agcaccacgt
     1261 ggacgcaaag ccttggtcta accagacctt cccaggactg acacctaagg atggagcagg
     1321 cacaagggtg agctacgcct tttggatcat ggccctgatc aatctgcctg tgccaatggc
     1381 cgtgctgatg ctgctgagca aggagcgcct gctgacctgc tgtccacaga ggcgccccct
     1441 gctgctgtcc gccgacgagc tggccctgga gacacagccc cctgagaagg aggatgcctc
     1501 ctctctgcca cccaagttcc agtctcacct gggccacgag gacctgttta gctgctgtca
     1561 gcggaagaac ctgagaggcg ccccttatag cttctttgcc atccacatca ccggcgccct
     1621 ggtgctgttc atgaccgatg gcctgacagg cgcctacagc gccttcgtgt actcttatgc
     1681 cgtggagaag ccactgtccg tgggacacaa ggtggcaggc tatctgccct ctctgttctg
     1741 gggctttatc accctgggcc ggctgctgtc catccccatc agctccagaa tgaagcctgc
     1801 caccatggtg ttcatcaacg tggtgggcgt ggtggtgaca ttcctggtgc tgctgatctt
     1861 ttcttacaat gtggtgttcc tgtttgtggg aaccgcaagc ctgggactgt tcctgtctag
     1921 cacctttccc tccatgctgg cctacacaga ggactctctg cagtataagg gctgcgcaac
     1981 cacagtgctg gtgacaggag caggagtggg agagatggtg ctgcagatgc tggtgggctc
     2041 catcttccag gcccagggct cctactcttt cctggtgtgc ggcgtgatct ttggctgtct
     2101 ggccttcacc ttttatatcc tgctgctgtt ctttcacaga atgcaccccg gcctgccttc
     2161 tgtgccaaca caggataggt ctatcggcat ggagaatagc gagtgttatc agcgcgaccc
     2221 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2281 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2341 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2401 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2461 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2521 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2581 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2641 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2701 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2761 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2821 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2881 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2941 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3001 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3061 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3121 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3181 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3241 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3301 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3361 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4081 tgctcacatg tt
//
