LOCUS       sequence-479349-        4041 bp    DNA     circular SYN 14-AUG-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 4041)
  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 4041)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 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-TMEM104_STOP
FEATURES             Location/Qualifiers
     source          1..4041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(44..130)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     193..210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     194..210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    226..325
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1823..1922)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1939..1958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1940..1958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1963..1979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1963..1979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2081..2100)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2092..2901
                     /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(2159..2178)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3048..3636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3537..3556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3790..3807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3966..3993)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
       61 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
      121 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
      181 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
      241 ttttgactga tagtgacctg ttcgttgcaa cacattgatg agcaatgctt ttttataatg
      301 ccaactttgt acaaaaaagc aggctccacc atggccggcg agatcacaga gaccggcgag
      361 ctgtacagct cctatgtggg cctggtgtac atgtttaacc tgatcgtggg aacaggcgcc
      421 ctgaccatgc caaaggcctt cgccaccgca ggatggctgg tgtccctggt gctgctggtg
      481 ttcctgggct ttatgtcttt cgtgaccaca acctttgtga tcgaggcaat ggcagcagca
      541 aacgcgcagt tgcattggaa acgcatggag aatctgaagg aggaggagga cgatgactct
      601 agcaccgcca gcgattccga cgtgctgatc agagataact acgagagggc cgagaagcgc
      661 cctatcctga gcgtgcagcg gagaggctcc cctaatccat ttgagatcac cgaccgggtg
      721 gagatgggcc agatggccag catgttcttt aacaaggtcg gcgtgaatct gttctacttt
      781 tgcatcatcg tgtacctgta tggcgatctg gccatctatg ccgccgccgt gccattctcc
      841 ctgatgcagg tgacatgctc tgccaccggc aacgatagct gtggcgtgga ggccgacaca
      901 aagtacaatg ataccgacag gtgttgggga ccactgagga gggtggatgc atacagaatc
      961 tatctggcca tcttcacact gctgctgggc ccttttacct tctttgacgt gcagaagaca
     1021 aagtatctgc agatcctgac cagcctgatg aggtggatcg ccttcgccgt gatgatcgtg
     1081 ctggccctga tccgcatcgg acacggacag ggagagggac acccacctct ggcagacttc
     1141 tctggcgtga ggaatctgtt tggcgtgtgc gtgtacagct tcatgtgcca gcactctctg
     1201 cccagcctga tcacacccgt gagctctaag cggcacctga ccagactggt gtttctggat
     1261 tacgtgctga tcctggcctt ctatggcctg ctgagcttta cagccatctt ctgctttcgc
     1321 ggcgattccc tgatggacat gtataccctg aacttcgcaa ggtgtgacgt ggtgggactg
     1381 gcagccgtgc gcttctttct gggcctgttt ccagtgttca caatctctac caattttccc
     1441 atcatcgccg tgacactgcg gaacaattgg aagaccctgt tccacagaga gggcggcaca
     1501 tacccttggg tggtggatag agtggtgttt ccaacaatca ccctggtgcc acccgtgctg
     1561 gtggccttct gcacccacga cctggagtcc ctggtgggaa tcacaggagc atacgcagga
     1621 accggaatcc agtatgtgat cccagccttt ctggtgtatc actgccggag agatacacag
     1681 ctggccttcg gctgtggcgt gtccaacaag cacaggtctc cctttcgcca caccttctgg
     1741 gtgggctttg tgctgctgtg ggccttctcc tgtttcatct ttgtgacagc caatatcatc
     1801 ctgtctgaga ccaagctgta agacccagct ttcttgtaca aagttggcat tataagaaag
     1861 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     1921 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1981 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     2041 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     2101 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     2161 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     2221 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     2281 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     2341 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     2401 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2461 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2521 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2581 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2641 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2701 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2761 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2821 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2881 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2941 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     3001 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3061 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3181 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3721 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3781 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3841 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3901 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3961 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     4021 gcgggcgtcc tgcccgccac c
//
