LOCUS       sequence-479351-        4026 bp    DNA     circular SYN 24-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 4026)
  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 4026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 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-LETM2_STOP
FEATURES             Location/Qualifiers
     source          1..4026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..231
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     465..482
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(641..668)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(760..846)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     895..917
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     909..926
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     910..926
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    942..1041
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2524..2623)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2640..2659)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2641..2659)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2664..2680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2664..2680)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2782..2801)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2793..3602
                     /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(2860..2879)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3749..4026,1..311)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
       61 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      121 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      181 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      241 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      301 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      361 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      421 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      481 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      541 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      601 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
      661 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
      721 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
      781 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
      841 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
      901 cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt
      961 gactgatagt gacctgttcg ttgcaacaca ttgatgagca atgctttttt ataatgccaa
     1021 ctttgtacaa aaaagcaggc tccaccatgg ccttctattc ctacaactct gtgctggcaa
     1081 tcgcaaggac caggttccca agccactttg tgcaccccac atgcagctcc tattccccat
     1141 cttgtgcctt tctgcacctg cccgactccc acctgaacaa gacctgcatg aagaattatg
     1201 agtctaagaa gtacagcgat ccttcccagc caggcaatac cgtgctgcac ccaggcacac
     1261 gcctgatcca gaagctgcac acctccacat gttggctgca ggaggtgcca ggcaagcctc
     1321 agctggagca ggcaaccaag cacccccagg tgacaagccc tcaggccacc aaggagacag
     1381 gcatggagat caaggagggc aagcagtcct atcggcagaa gatcatggac gagctgaagt
     1441 actattacaa cggcttctac ctgctgtgga tcgatgcaaa ggtggcagca aggatggtgt
     1501 ggaggctgct gcacggacag gtgctgacca ggagagagag gcgccggctg ctgagaacat
     1561 gcgtggactt ctttaggctg gtgcctttca tggtgtttct gatcgtgccc ttcatggagt
     1621 ttctgctgcc cgtgttcctg aagctgtttc ctgagatgct gccaagcacc ttcgagagcg
     1681 agtccaagaa ggaggagaag cagaagaaga agatggccgt gaagctggag ctggccaagt
     1741 ttctgcagga gaccatgaca gagatggcca gaaggaatag agccaagatg ggcgatgcct
     1801 ccacacagct gtctagctat gtgaagcagg tgcagaccgg ccacaagccc tctacaaagg
     1861 agatcgtgag attcagcaag ctgtttgagg accagctggc cctggagcac ctggataggc
     1921 ctcagctggt ggccctgtgc aagctgctgg agctgcagac cttcggcaca aacaatctgc
     1981 tgaggtttca gctgctgatg aagctgaagt ccatcaaggc cgacgatgag atcatcgcca
     2041 aggagggcgt gaccgccctg tctgtgagcg agctgcaggc agcatgtaga gcaaggggaa
     2101 tgcgctctct gggactgacc gaggagcagc tgaggcagca gctgacagag tggcaggacc
     2161 tgcacctgaa ggagaacgtg cccccttccc tgctgctgct gtctcggacc ttctacctga
     2221 tcgatgtgaa gccaaagccc atcgagatcc ctctgtctgg cgaggcccca aagacagaca
     2281 tcctggtgga gctgcctacc ttcacagaga gcaaggagaa tatggtggat ctggccccac
     2341 agctgaaggg caccaaggac gaggacttca tccagccacc ccctgtgaca tcctctccaa
     2401 tcaccccctc tacacctatc agcctgccaa agggaccaat caccagctcc gaggagccca
     2461 cactgcaggc caagagccag atgaccgccc agaactccaa ggcctctagc aagggcgcct
     2521 gagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     2581 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     2641 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     2701 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     2761 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     2821 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     2881 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     2941 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     3001 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     3061 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     3121 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3181 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     3241 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3301 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3361 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3421 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3481 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3541 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3601 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     3661 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3721 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3841 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4021 tggact
//
