LOCUS       sequence-261102-        4941 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 4941)
  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 4941)
  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_SLC6A5
FEATURES             Location/Qualifiers
     source          1..4941
                     /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(3066..3165)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3182..3201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3183..3201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3206..3222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3206..3222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3324..3343)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3335..4144
                     /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(3402..3421)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4291..4879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4780..4799
                     /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 accatggact gctccgcccc caaggagatg aacaagctgc cagccaattc
      721 tccagaggca gcagcagcac agggacaccc agatggacct tgtgcaccac ggacctctcc
      781 agagcaggag ctgcctgcag cagcagcacc acctccacca cgggtgccta gatccgcctc
      841 tacaggcgcc cagaccttcc agagcgccga cgcaagggca tgcgaggccg agagaccagg
      901 cgtgggcagc tgtaagctga gctccccaag agcacaggca gcatccgccg ccctgaggga
      961 tctgcgcgag gcccagggcg cccaggcctc ccctccaccc ggctctagcg gaccaggaaa
     1021 cgcactgcac tgcaagatcc catttctgag gggaccagag ggcgacgcaa acgtgagcgt
     1081 gggcaagggc acactggagc gcaacaatac ccctgtggtg ggctgggtga acatgagcca
     1141 gtccaccgtg gtgctggcca cagatggcat cacctctgtg ctgccaggaa gcgtggcaac
     1201 cgtggcaaca caggaggacg agcagggcga tgagaacaag gcccggggca attggtcctc
     1261 taagctggac ttcatcctgt ccatggtggg atacgcagtg ggactgggaa acgtgtggag
     1321 attcccatat ctggcctttc agaatggcgg cggcgccttc ctgatccctt acctgatgat
     1381 gctggccctg gcaggactgc caatcttctt tctggaggtg agcctgggac agtttgcatc
     1441 ccagggaccc gtgagcgtgt ggaaggcaat ccctgccctg cagggatgcg gaatcgcaat
     1501 gctgatcatc tccgtgctga tcgccatcta ctataacgtg atcatctgtt acaccctgtt
     1561 ctatctgttt gcctctttcg tgagcgtgct gccctggggc agctgcaaca atccttggaa
     1621 tacaccagag tgtaaggata agaccaagct gctgctggac agctgcgtga tctccgatca
     1681 cccaaagatc cagatcaaga actccacatt ctgtatgacc gcctacccca acgtgacaat
     1741 ggtgaatttc acctctcagg ccaacaagac cttcgtgagc ggcagcgagg agtactttaa
     1801 gtatttcgtg ctgaagatca gcgccggcat cgagtaccct ggagagatca ggtggccact
     1861 ggccctgtgc ctgttcctgg cctgggtcat cgtgtacgcc tctctggcca agggcatcaa
     1921 gaccagcggc aaggtggtgt acttcaccgc cacatttccc tatgtggtgc tggtcatcct
     1981 gctgatcaga ggagtgacac tgcctggagc aggagcagga atctggtact ttatcacccc
     2041 aaagtgggag aagctgaccg acgccacagt gtggaaggat gccgccacac agatcttctt
     2101 ttccctgtct gccgcatggg gaggactgat caccctgagc tcctacaaca agttccacaa
     2161 caattgctat agggacacac tgatcgtgac ctgtacaaat tccgccacct ctatctttgc
     2221 cggcttcgtg atcttttccg tgatcggctt tatggccaac gagaggaagg tgaacatcga
     2281 gaatgtggcc gatcagggcc caggcatcgc cttcgtggtg tatcctgagg ccctgaccag
     2341 gctgccactg agcccttttt gggccatcat cttctttctg atgctgctga cactgggcct
     2401 ggacaccatg ttcgccacca tcgagacaat cgtgaccagc atctccgatg agttccctaa
     2461 gtacctgcgg acacacaagc cagtgtttac cctgggctgc tgtatctgct tctttatcat
     2521 gggcttccct atgatcacac agggcggcat ctatatgttt cagctggtgg acacctacgc
     2581 cgcctcttat gccctggtca tcatcgccat cttcgagctg gtgggcatca gctacgtgta
     2641 tggcctgcag aggttttgtg aggatatcga gatgatgatc ggcttccagc ccaacatctt
     2701 ttggaaggtg tgctgggcct tcgtgacccc tacaatcctg accttcatcc tgtgcttcag
     2761 cttttaccag tgggagccca tgacctatgg cagctaccgc tatcctaatt ggtccatggt
     2821 gctgggctgg ctgatgctgg cctgcagcgt gatctggatc cccatcatgt tcgtgatcaa
     2881 gatgcacctg gcccctggca ggtttatcga gcgcctgaag ctggtgtgct cccctcagcc
     2941 agactgggga cccttcctgg cacagcacag gggagagaga tacaagaata tgatcgaccc
     3001 actgggcaca tctagcctgg gcctgaagct gcccgtgaag gatctggagc tgggcaccca
     3061 gtgcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     3121 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3181 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3241 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     3301 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     3361 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     3421 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     3481 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3541 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3601 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3661 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3721 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3781 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3841 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3901 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3961 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4021 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4081 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4141 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4201 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     4261 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4321 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4381 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4441 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4501 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4561 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4621 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4681 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4741 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4801 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4861 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4921 ctggcctttt gctcacatgt t
//
