LOCUS       sequence-259257-        4128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  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 4128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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_FLVCR2
FEATURES             Location/Qualifiers
     source          1..4128
                     /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(2253..2352)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2369..2388)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2370..2388)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2393..2409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2393..2409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2511..2530)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2522..3331
                     /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(2589..2608)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3478..4066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3967..3986
                     /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 accatggtga acgagggccc taatcaggag gagtctgacg atacccctgt
      721 gccagagagc gccctgcagg cagacccatc tgtgagcgtg cacccttccg tgtctgtgca
      781 cccaagcgtg tccatcaacc cctctgtgag cgtccatcct agctcctctg cccacccatc
      841 cgccctggca cagccatctg gactggcaca ccctagctcc tctggaccag aggatctgtc
      901 tgtgatcaag gtgagccgga gaaggtgggc cgtggtgctg gtgttcagct gctactccat
      961 gtgcaattcc tttcagtgga tccagtatgg ctctatcaac aatatcttca tgcactttta
     1021 cggcgtgagc gccttcgcca tcgactggct gtccatgtgc tacatgctga catatatccc
     1081 actgctgctg cctgtggcat ggctgctgga gaagtttggc ctgcgcacca tcgccctgac
     1141 aggatctgcc ctgaactgcc tgggagcatg ggtgaagctg ggatccctga agccacacct
     1201 gttccccgtg accgtggtgg gccagctgat ctgtagcgtg gcccaggtgt tcatcctggg
     1261 catgccatcc agaatcgcca gcgtgtggtt tggcgccaac gaggtgagca cagcctgttc
     1321 cgtggccgtg ttcggcaatc agctgggcat cgccatcggc tttctggtgc cccctgtgct
     1381 ggtgcccaat atcgaggaca gggatgagct ggcctaccac atcagcatca tgttctatat
     1441 catcggagga gtggccaccc tgctgctgat cctggtcatc atcgtgttta aggagaagcc
     1501 caagtatcca ccatccaggg cacagtccct gtcttacgca ctgacatctc ctgatgccag
     1561 ctatctgggc tccatcgccc ggctgttcaa gaacctgaat tttgtgctgc tggtcatcac
     1621 ctacggcctg aacgccggcg ccttttatgc cctgtccaca ctgctgaata gaatggtcat
     1681 ctggcactat ccaggagagg aggtgaacgc aggaaggatc ggcctgacca tcgtgatcgc
     1741 aggaatgctg ggagccgtga tcagcggcat ctggctggac cggtccaaga cctacaagga
     1801 gaccacactg gtggtgtata tcatgacact ggtgggcatg gtggtgtaca ccttcacact
     1861 gaacctgggc cacctgtggg tggtgtttat caccgccggc acaatgggct tctttatgac
     1921 cggctatctg cctctgggct tcgagtttgc cgtggagctg acatacccag agtctgaggg
     1981 catcagctcc ggcctgctga atatcagcgc ccaggtgttc ggcatcatct ttaccatctc
     2041 ccagggccag atcatcgaca actacggcac aaagcctggc aatatcttcc tgtgcgtgtt
     2101 tctgaccctg ggagccgccc tgacagcctt catcaaggcc gatctgcgcc ggcagaaggc
     2161 caacaaggag accctggaga ataagctgca ggaggaggag gaggagtcta ataccagcaa
     2221 ggtgccaaca gccgtgagcg aggatcacct ggacccagct ttcttgtaca aagttggcat
     2281 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     2341 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     2401 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2461 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2521 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2581 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2641 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2701 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2761 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2821 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2881 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2941 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     3001 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     3061 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     3121 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     3181 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     3241 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     3301 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     3361 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3421 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
