LOCUS       sequence-479348-        3123 bp    DNA     circular SYN 23-JUL-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 3123)
  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 3123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 23, 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-SLC31A1_STOP
FEATURES             Location/Qualifiers
     source          1..3123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(132..231)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(247..264)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(247..263)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(256..278)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      327..413
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      505..532
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(691..708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(862..1450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(942..961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1597..2406)
                     /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     2320..2339
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2398..2417
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2519..2535
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2519..2535
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2540..2559
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2540..2558
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2576..2675
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 catcatcatg gagctgtcgc ctcctccgtg gctgtgagag gcggatgtgg tagggtggtg
       61 gtggctgggc tgcatggtgc tgttagaatc catgtaggac atgcccatgt ggtggctgtg
      121 gtccatggtg gagcctgctt ttttgtacaa agttggcatt ataaaaaagc attgctcatc
      181 aatgtgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggggcccgag
      241 cttaagactg gccgtcgttt tacaacgtcg tgactgggaa aacatccatg ctagcgttaa
      301 cgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg
      361 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      421 gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
      481 aaactgccag gcatcaaact aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      541 tacaaactct tcctggctag cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt
      601 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      661 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      721 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      781 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      841 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1261 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1441 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc
     1501 gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat
     1561 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     1621 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1681 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1741 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1801 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1861 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1921 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     1981 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2041 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2101 ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2161 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2221 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2281 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2341 atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgtttccc gttgaatatg
     2401 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     2461 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcca
     2521 ggaaacagct atgaccatgt aatacgactc actatagggg atatcagctg gatggcaaat
     2581 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt
     2641 tcttataatg ccaactttgt acaagaaagc tgggtcttag tgacagtgct ctgtgatatc
     2701 caccaccacg gccttcttcc aagagaacag aaaatagcct gttcctgctc ctgctgccac
     2761 tgcgatgcac agatagccat tgtaggtcat gaagatcagc atcagaaagt aagagatgac
     2821 cacctggatg atgtgcagca ctgtctgcag caggtgggga aaggacagca tctgctggcc
     2881 cactgtcttg tgggtctcca tcaggatggt gccatttggg ccaggcacgg gcatgctgtt
     2941 ataccggata gacacctggg actttctcag cagagactcc cgggcgatct tcaggccctc
     3001 gtagaacatg gccagcagga acacggccac aaaggctcct gccatctctc ctgctgtgtt
     3061 gatgaccagg ccggaaaaca gcagctccac gttcttgaag ccaaaataga aggtcattgg
     3121 cat
//
