LOCUS       sequence-259571-        3861 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 3861)
  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 3861)
  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_SLC10A4
FEATURES             Location/Qualifiers
     source          1..3861
                     /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(1986..2085)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2102..2121)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2103..2121)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2126..2142)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2126..2142)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2244..2263)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2255..3064
                     /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(2322..2341)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3211..3799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3700..3719
                     /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 accatggacg gcaacgataa tgtgacactg ctgttcgccc ccctgctgag
      721 ggacaactac accctggcac caaatgcaag ctccctggga cctggaacag atctggccct
      781 ggcacctgca tctagcgccg gacctggccc aggactgtcc ctgggaccag gaccttcttt
      841 cggctttagc ccaggaccaa cccctacacc agagccaacc acatctggac tggcaggagg
      901 agcagcatcc cacggacctt ctccattccc aagaccttgg gcaccacacg cactgccctt
      961 ttgggacacc cccctgaacc acggcctgaa cgtgttcgtg ggagccgccc tgtgcatcac
     1021 aatgctgggc ctgggctgta ccgtggatgt gaaccacttc ggagcacacg tgcggagacc
     1081 tgtgggcgcc ctgctggccg ccctgtgcca gtttggactg ctgccactgc tggccttcct
     1141 gctggccctg gcctttaagc tggacgaggt ggcagcagtg gccgtgctgc tgtgcggatg
     1201 ctgtcctggc ggcaacctga gcaatctgat gtccctgctg gtggacggcg atatgaacct
     1261 gagcatcatc atgacaatct cctctaccct gctggccctg gtgctgatgc ctctgtgcct
     1321 gtggatctat tcctgggcct ggatcaatac ccctatcgtg cagctgctgc cactgggcac
     1381 cgtgaccctg accctgtgca gcacactgat cccaatcggc ctgggcgtgt tcatcaggta
     1441 caagtattcc cgcgtggccg attacatcgt gaaggtgagc ctgtggtccc tgctggtgac
     1501 cctggtggtg ctgtttatca tgacaggaac catgctggga ccagagctgc tggcatctat
     1561 cccagcagcc gtgtatgtga tcgccatctt catgccactg gcaggatacg caagcggata
     1621 tggcctggcc accctgtttc acctgccccc taactgcaag cggaccgtgt gcctggagac
     1681 cggctctcag aatgtgcagc tgtgcacagc catcctgaag ctggccttcc caccccagtt
     1741 tatcggctct atgtacatgt tcccactgct gtatgccctg tttcagagcg ccgaggccgg
     1801 catcttcgtg ctgatctaca agatgtatgg ctccgagatg ctgcacaaga gagaccccct
     1861 ggacgaggat gaggacaccg atatctccta caagaagctg aaggaggagg agatggccga
     1921 tacatcttat ggcaccgtga aggccgagaa tatcatcatg atggagacag cccagaccag
     1981 cctggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2041 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2101 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2161 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2221 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2281 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2341 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2401 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2461 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2521 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2581 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2641 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2701 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2761 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2821 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2881 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2941 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3001 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3061 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3121 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3841 ctggcctttt gctcacatgt t
//
