LOCUS       sequence-265826-        3774 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 3774)
  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 3774)
  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_SLC35F1
FEATURES             Location/Qualifiers
     source          1..3774
                     /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(1899..1998)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2015..2034)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2016..2034)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2039..2055)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2039..2055)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2157..2176)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2168..2977
                     /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(2235..2254)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3124..3712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3613..3632
                     /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 accatgatcc cccctgagca gcctcagcag cagctgcagc caccatctcc
      721 agcacctcca aaccacgtgg tgaccacaat cgagaatctg cctgcagagg gatccggagg
      781 aggaggatcc ctgtctgcca gctccagggc aggcgtgcgc cagaggatca gaaaggtgct
      841 gaaccgcgag atgctgatct ctgtggccct gggacaggtg ctgagcctgc tgatctgcgg
      901 catcggcctg acatctaagt acctgagcga ggactttcac gccaacaccc cagtgttcca
      961 gagctttctg aattacatcc tgctgttcct ggtgtatacc acaaccctgg ccgtgaggca
     1021 gggagaggag aacctgctgg ccatcctgcg gagaaggtgg tggaagtaca tgatcctggg
     1081 cctgatcgac ctggaggcca attatctggt ggtgaaggcc taccagtata caaccctgac
     1141 atccatccag ctgctggatt gttttgtgat ccccgtggtc atcctgctgt cttggttctt
     1201 tctgctgatc cggtataagg ccgtgcactt catcggcatc gtggtgtgca tcctgggcat
     1261 gggctgtatg gtgggcgccg acgtgctggt gggcagacac cagggagcag gagagaacaa
     1321 gctggtgggc gatctgctgg tgctgggagg agccacactg tacggcatct ctaacgtgtg
     1381 ggaggagtat atcatcagga ccctgagccg cgtggagttt ctgggcatga tcggcctgtt
     1441 cggcgccttc ttttccggca tccagctggc catcatggag cacaaggagc tgctgaaggt
     1501 gccctgggac tggcagatcg gcctgctgta cgtgggcttc tccgcctgca tgtttggcct
     1561 gtattctttc atgcctgtgg tcatcaagaa gaccagcgcc accagcgtga acctgagcct
     1621 gctgacagcc gatctgtact ccctgttttg tggcctgttc ctgtttcact acaagttcag
     1681 cggcctgtat ctgctgtcct tctttaccat cctgatcggc ctggtgctgt actctagcac
     1741 aagcacctat atcgcccagg acccacgggt gtacaagcag ttcagaaatc cctccggccc
     1801 tgtggtggat ctgccaacaa ccgcccaggt ggagccctcc gtgacatata cctctctggg
     1861 acaggagacc gaggaggagc cacacgtgcg ggtggccgac ccagctttct tgtacaaagt
     1921 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1981 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2041 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2101 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2161 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2221 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2281 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2341 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2401 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2461 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2521 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2581 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2641 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2701 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2761 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2821 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2881 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2941 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3001 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3061 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3241 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3721 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
