LOCUS       sequence-261098-        4041 bp    DNA     circular SYN 14-APR-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 4041)
  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 4041)
  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_SLC19A2
FEATURES             Location/Qualifiers
     source          1..4041
                     /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(2166..2265)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2282..2301)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2283..2301)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2306..2322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2306..2322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2424..2443)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2435..3244
                     /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(2502..2521)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3391..3979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3880..3899
                     /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 accatggacg tgccaggccc cgtgtccagg agagcagcag cagcagcagc
      721 aaccgtgctg ctgcggacag ccagagtgag gagggagtgc tggttcctgc ctaccgccct
      781 gctgtgcgca tacggcttct ttgcctctct gcggcccagc gagccttttc tgaccccata
      841 tctgctgggc cccgacaaga acctgacaga gagagaggtg ttcaatgaga tctaccccgt
      901 gtggacctac tcttatctgg tgctgctgtt ccccgtgttc ctggccacag attacctgcg
      961 gtataagcca gtggtgctgc tgcagggcct gagcctgatc gtgacctggt ttatgctgct
     1021 gtacgcccag ggcctgctgg ccatccagtt cctggagttc ttttatggca tcgccaccgc
     1081 cacagagatc gcctactatt cctacatcta ttctgtggtg gatctgggca tgtaccagaa
     1141 ggtgaccagc tattgccggt ccgccaccct ggtgggcttt acagtgggct ctgtgctggg
     1201 acagatcctg gtgagcgtgg caggatggtc cctgttctct ctgaacgtga tctccctgac
     1261 ctgcgtgtct gtggccttcg ccgtggcctg gtttctgcct atgccacaga agtccctgtt
     1321 ctttcaccac atcccatcta catgtcagag agtgaacggc atcaaggtgc agaatggcgg
     1381 catcgtgacc gacacacctg ccagcaacca cctgccaggc tgggaggata tcgagtccaa
     1441 gatccccctg aatatggagg agccacctgt ggaggagcca gagcctaagc cagacaggct
     1501 gctggtgctg aaggtgctgt ggaatgattt tctgatgtgc tacagctcca ggccactgct
     1561 gtgctggagc gtgtggtggg cactgtccac ctgcggctac ttccaggtgg tgaactatac
     1621 acagggcctg tgggagaaag tgatgccttc tagatacgcc gccatctata atggcggcgt
     1681 ggaggccgtg agcaccctgc tgggagcagt ggccgtgttt gcagtgggct acatcaagat
     1741 ctcttggagc acctggggcg agatgacact gagcctgttc tccctgctga tcgcagcagc
     1801 cgtgtatatc atggacacag tgggcaacat ctgggtgtgc tacgccagct atgtggtgtt
     1861 taggatcatc tacatgctgc tgatcaccat cgccacattc cagatcgccg ccaacctgtc
     1921 catggagcgg tacgccctgg tgttcggcgt gaataccttt atcgccctgg ccctgcagac
     1981 cctgctgaca ctgatcgtgg tggacgcatc cggactggga ctggagatca ccacacagtt
     2041 cctgatctac gcctcttatt ttgccctgat cgcagtggtg ttcctggcat ctggagccgt
     2101 gagcgtgatg aagaagtgtc gcaagctgga ggatccccag tctagctccc aggtgaccac
     2161 aagcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2221 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2281 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2341 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2401 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2461 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2521 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2581 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2641 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2701 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2761 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2821 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2881 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2941 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3001 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3061 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3121 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3181 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     3241 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3301 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3361 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3421 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3481 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3841 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3901 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3961 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4021 ctggcctttt gctcacatgt t
//
