LOCUS       sequence-444347-        4014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4014)
  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 4014)
  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-SLC16A4_STOP
FEATURES             Location/Qualifiers
     source          1..4014
                     /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(2139..2238)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2255..2274)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2256..2274)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2279..2295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2279..2295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2397..2416)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2408..3217
                     /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(2475..2494)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3364..3952
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3853..3872
                     /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 accatgctga agcgggaggg caaggtgcag ccatacacca agacactgga
      721 cggcggctgg ggctggatga tcgtgatcca cttctttctg gtgaacgtgt ttgtgatggg
      781 catgaccaag acattcgcca tcttctttgt ggtgtttcag gaggagttcg agggcaccag
      841 cgagcagatc ggctggatcg gctccatcat gagctccctg agattctgcg caggaccact
      901 ggtggcaatc atctgtgata tcctgggcga gaagaccaca agcatcctgg gcgcctttgt
      961 ggtgacaggc ggctacctga tctctagctg ggccaccagc atccccttcc tgtgcgtgac
     1021 aatgggactg ctgcctggac tgggatccgc ctttctgtac caggtggccg ccgtggtgac
     1081 cacaaagtat ttcaagaagc ggctggccct gtccaccgca atcgcaagat ctggcatggg
     1141 cctgaccttt ctgctggccc cctttacaaa gttcctgatc gacctgtatg attggacagg
     1201 cgccctgatc ctgttcggag caatcgccct gaatctggtg ccttcctcta tgctgctgag
     1261 gccaatccac atcaagagcg agaacaattc cggcatcaag gacaagggaa gctccctgtc
     1321 tgcccacgga cctgaggcac acgcaaccga gacacactgc cacgagaccg aggagtctac
     1381 aatcaaggat agcaccacac agaaggccgg cctgccatcc aagaacctga ccgtgtctca
     1441 gaatcagagc gaggagttct acaacggccc caaccggaat agactgctgc tgaagtccga
     1501 cgaggagtct gataaagtga tctcctggtc ttgtaagcag ctgtttgaca tcagcctgtt
     1561 caggaatcct ttcttttaca tctttacctg gagcttcctg ctgtcccagc tggcctactt
     1621 catcccaacc ttccacctgg tggcaagggc aaagacactg ggcatcgaca tcatggatgc
     1681 ctcttacctg gtgagcgtgg caggaatcct ggagaccgtg tcccagatca tctctggctg
     1741 ggtggccgat cagaactgga tcaagaagta ccactatcac aagagctatc tgatcctgtg
     1801 cggcatcaca aatctgctgg ccccactggc caccacattt cccctgctga tgacctacac
     1861 aatctgcttt gccatcttcg caggaggata tctggccctg atcctgcctg tgctggtgga
     1921 cctgtgcagg aactccaccg tgaatcgctt cctgggcctg gcctctttct ttgcaggaat
     1981 ggccgtgctg agcggaccac ctatcgccgg ctggctgtac gattataccc agacatacaa
     2041 cggcagcttt tatttctccg gcatctgtta tctgctgtct agcgtgtcct tctttttcgt
     2101 gcccctggcc gagcggtgga agaattctct gacctaagac ccagctttct tgtacaaagt
     2161 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2221 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2281 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2341 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2401 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2461 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2521 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2581 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2641 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2701 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2761 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2821 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2881 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2941 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3001 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3061 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3121 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3181 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3241 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3301 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3361 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3421 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3481 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3541 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3601 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3661 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3721 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3781 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3841 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3901 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3961 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
