LOCUS       sequence-436371-        4857 bp    DNA     circular SYN 14-APR-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 4857)
  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 4857)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-SLC45A4_STOP
FEATURES             Location/Qualifiers
     source          1..4857
                     /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(2982..3081)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3098..3117)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3099..3117)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3122..3138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3122..3138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3240..3259)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3251..4060
                     /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(3318..3337)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4207..4795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4696..4715
                     /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 accatgaaga tggctcccca gaacgctgat cccgagtcta tgcaggtcca
      721 ggaactgtcc gtcccactgc ccgaccccca gaaagcagga ggcgccgagg ccgagaactg
      781 cgagaccatc tctgagggct ccatcgacag aatccccatg agactgtggg taatgcacgg
      841 cgccgtgatg ttcggccgcg agttttgtta cgccatggag accgccctgg tgacccctat
      901 cctgctgcag atcggcctgc cagagcagta ttactccctg acctggttcc tgtccccaat
      961 cctgggcctg atcttcacac ccctgatcgg cagcgcctct gataggtgca cactgtcttg
     1021 gggcagacgg agacctttca tcctggccct gtgcgtgggc gtgctgtttg gcgtggccct
     1081 gtttctgaat ggctctgcca tcggcctggc cctgggcgac gtgcccaaca gacagcctat
     1141 cggcatcgtg ctgacagtgc tgggcgtggt ggtgctggac ttttccgccg atgccacaga
     1201 gggcccaatc agagcctacc tgctggacgt ggtggattcc gaggagcagg acatggccct
     1261 gaatatccac gcctttagcg ccggcctggg cggcgccatc ggctatgtgc tgggcggcct
     1321 ggattggacc cagacatttc tgggctcttg gtttagaacc cagaatcagg tgctgttctt
     1381 cttcgccgcc atcatcttca cagtgtccgt ggccctgcac ctgttttcca tcgatgagga
     1441 gcagtattcc cctcagcagg agagatctgc cgaggagccc ggcgccctgg atggcggcga
     1501 gccccacggc gtgcctgcct ttcctgacga ggtgcagagc gagcacgagc tggccctgga
     1561 ctatccagat gtggacatca tgaggtccaa gtccgatagc gccctgcacg tgcccgatac
     1621 agccctggat ctggagccag agctgctgtt cctgcacgat atcgagccat ctatcttcca
     1681 cgatgcctcc taccccgcca ccccaagaag cacctctcag gagctggcca agaccaagct
     1741 gccaaggctg gccacctttc tgaaggaggc cgccaaggag gatgagacac tgctggacaa
     1801 ccacctgaac gaggccaagg tgccaaacgg cagcggctct ccaaccaagg atgccctggg
     1861 cggctatacc cgggtggata ccaagccatc tgccacatct agctccatga gacggagaag
     1921 acacgccttc agacggcagg cctcctctac attcagctat tacggcaagc tgggctccca
     1981 ctgctataga taccggagag ccaatgccgt ggtgctgatc aagccttcca ggagcatgtc
     2041 cgacctgtat gacatgcaga agagacagag acagcacagg cacagaaacc agtctggcgc
     2101 caccacatcc agcggcgaca ccgagagcga ggagggcgag ggcgagacaa ccgtgagact
     2161 gctgtggctg tctatgctga agatgccaag agagctgatg aggctgtgcc tgtgccacct
     2221 gctgacctgg tttagcgtga tcgccgaggc cgtgttttat accgacttta tgggccaggt
     2281 aatcttcgag ggcgatccta aggccccaag caattccacc gcctggcagg cctataacgc
     2341 cggcgtgaag atgggctgct ggggcctggt aatctatgcc gccacaggcg ccatctgttc
     2401 cgccctgctg cagaagtacc tggacaatta tgatctgagc gtgagagtga tctacgtgct
     2461 gggcacactg ggctttagcg tgggcaccgc cgtgatggcc atgttcccca acgtgtacgt
     2521 ggccatggtg accatcagca caatgggcat cgtgagcatg tccatcagct actgcccata
     2581 tgccctgctg ggccagtatc acgacatcaa gcagtatatc caccactctc ccggcaatag
     2641 caagcggggc tttggcatcg attgcgccat cctgtcctgt caggtgtaca tctcccagat
     2701 cctggtggcc tccgccctgg gcggcgtggt ggatgccgtg ggcaccgtga gagtgatccc
     2761 tatggtggcc tccgtgggct cttttctggg cttcctgacc gccacattcc tggtaatcta
     2821 ccctaatgtg agcgaggagg ccaaggagga gcagaagggc ctgtcttccc ctctggccgg
     2881 cgagggcagg gccggcggca actctgagaa gcccacagtg ctgaaactga caagaaagga
     2941 agggctgcag ggacccgtgg aaacagaatc cgtcgtctaa gacccagctt tcttgtacaa
     3001 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     3061 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     3121 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3181 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     3241 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
     3301 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     3361 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     3421 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     3481 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     3541 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     3601 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     3661 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     3721 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     3781 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     3841 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     3901 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     3961 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     4021 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     4081 gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     4141 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4201 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4261 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4321 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4381 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4441 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4501 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4561 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4621 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4681 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4741 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4801 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
