LOCUS       sequence-345627-        4257 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 4257)
  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 4257)
  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-SLC13A5_STOP
FEATURES             Location/Qualifiers
     source          1..4257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1814..1913)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1930..1949)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1931..1949)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1954..1970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1954..1970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2072..2091)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2083..2892
                     /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(2150..2169)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3039..3627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3528..3547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3781..3798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3957..3984)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4076..4162)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4211..4233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4225..4242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4226..4242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc ctctgccctg
      121 agctacgtgt ccaagtttaa gtccttcgtg atcctgtttg tgaccccact gctgctgctg
      181 cccctggtca tcctgatgcc agccaagttc gtgaggtgcg cctacgtgat catcctgatg
      241 gccatctatt ggtgtaccga agtgatcccc ctggccgtga caagcctgat gcctgtgctg
      301 ctgtttccac tgttccagat cctggactcc cgccaggtgt gcgtgcagta tatgaaggat
      361 accaacatgc tgtttctggg aggactgatc gtggcagtgg cagtggagag gtggaatctg
      421 cacaagagga tcgccctgcg cacactgctg tgggtgggag caaagcctgc aaggctgatg
      481 ctgggcttca tgggcgtgac cgccctgctg tctatgtgga tcagcaacac agccaccaca
      541 gccatgatgg tgccaatcgt ggaggccatc ctgcagcaga tggaggccac atccgccgca
      601 accgaggcag gactggagct ggtggacaag ggcaaggcca aggagctgcc aggctctcag
      661 gtcatcttcg agggacccac cctgggacag caggaggatc aggagcggaa gcggctgtgc
      721 aaggccatga cactgtgcat ctgttacgca gcatccatcg gaggaacagc caccctgaca
      781 ggaaccggac ctaacgtggt gctgctggga cagatgaatg agctgtttcc agacagcaag
      841 gatctggtga acttcgcctc ctggttcgcc tttgccttcc caaatatgct ggtcatgctg
      901 ctgtttgcct ggctgtggct gcagttcgtg tacatgcggt ttaacttcaa gaagtcctgg
      961 ggctgcggcc tggagtctaa gaagaatgag aaggccgccc tgaaggtgct gcaggaggag
     1021 tatcggaagc tgggccccct gtctttcgcc gagatcaacg tgctgatctg tttctttctg
     1081 ctggtcatcc tgtggtttag cagagaccct ggcttcatgc caggatggct gaccgtggca
     1141 tgggtggagg gagagacaaa gtacgtgagc gatgccaccg tggccatctt tgtggccaca
     1201 ctgctgttca tcgtgccctc tcagaagcct aagtttaatt tccggagcca gacagaggag
     1261 gagagaaaga ccccctttta tccccctcca ctgctggact ggaaggtgac ccaggagaag
     1321 gtgccttggg gcatcgtgct gctgctggga ggaggattcg cactggcaaa gggatctgag
     1381 gcaagcggac tgtccgtgtg gatgggcaag cagatggagc ctctgcacgc agtgccacct
     1441 gcagccatca ccctgatcct gtccctgctg gtggccgtgt ttacagagtg cacctctaac
     1501 gtggccacca caaccctgtt tctgcccatc ttcgccagca tgtcccggtc tatcggcctg
     1561 aatcccctgt acatcatgct gccttgtaca ctgagcgcct cctttgcctt catgctgcct
     1621 gtggccaccc cacccaacgc catcgtgttc acatatggcc acctgaaggt ggccgacatg
     1681 gtgaagaccg gcgtgatcat gaacatcatc ggcgtgtttt gcgtgttcct ggccgtgaat
     1741 acatggggca gagccatctt tgacctggat cacttccccg attgggccaa tgtgacacac
     1801 atcgagacct aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1861 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1921 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1981 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2041 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2101 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2161 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2221 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2281 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2341 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2401 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2461 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2521 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2581 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2641 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2701 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2761 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2821 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2881 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2941 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3001 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3061 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3841 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3901 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3961 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4021 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4081 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4141 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4201 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
