LOCUS       sequence-261170-        4302 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 4302)
  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 4302)
  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_SLC8B1
FEATURES             Location/Qualifiers
     source          1..4302
                     /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(2427..2526)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2543..2562)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2544..2562)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2567..2583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2567..2583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2685..2704)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2696..3505
                     /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(2763..2782)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3652..4240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4141..4160
                     /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 accatggcag gccggagact gaacctgaga tgggccctgt ctgtgctgtg
      721 cgtgctgctg atggcagaga ccgtgagcgg aacaagggga agctccaccg gagcacacat
      781 ctctccccag tttcctgcca gcggcgtgaa tcagacaccc gtggtggact gcagaaaggt
      841 gtgcggcctg aacgtgtccg acaggtgcga tttcatccgc accaatcctg actgtcactc
      901 tgatggcggc tacctggatt atctggaggg catcttctgc cactttccac ctagcctgct
      961 gcctctggca gtgaccctgt acgtgtcctg gctgctgtat ctgtttctga tcctgggcgt
     1021 gacagccgcc aagttctttt gtccaaacct gtccgccatc tctaccacac tgaagctgag
     1081 ccacaacgtg gccggcgtga ccttcctggc ctttggcaat ggcgccccag acatcttctc
     1141 cgccctggtg gccttttctg atccacacac agcaggactg gccctgggcg ccctgttcgg
     1201 agcaggcgtg ctggtgacca cagtggtggc aggaggaatc accatcctgc accctttcat
     1261 ggcagcatcc aggccattct ttagagacat cgtgttttac atggtggccg tgttcctgac
     1321 ctttctgatg ctgttcaggg gaagggtgac actggcatgg gcactgggat acctgggact
     1381 gtacgtgttt tatgtggtga ccgtgatcct gtgcacatgg atctatcaga ggcagaggcg
     1441 cggctctctg ttctgtccta tgccagtgac ccccgagatc ctgtccgact ctgaggagga
     1501 tcgggtgtct agcaacacaa atagctacga ctatggcgat gagtacagac ctctgttctt
     1561 ttatcaggag accacagccc agatcctggt gagggccctg aatccactgg actacatgaa
     1621 gtggcggaga aagagcgcct attggaaggc cctgaaggtg ttcaagctgc ccgtggagtt
     1681 tctgctgctg ctgaccgtcc cagtggtgga ccccgataag gacgatcaga actggaagcg
     1741 ccccctgaat tgtctgcacc tggtcatctc ccctctggtc gtggtgctga ccctgcagtc
     1801 tggcacatac ggcgtgtatg agatcggagg actggtgccc gtgtgggtgg tggtggtcat
     1861 cgcaggaaca gccctggcat ccgtgacctt ctttgccaca agcgattccc agccaccaag
     1921 gctgcactgg ctgttcgcct ttctgggctt tctgacctct gccctgtgga tcaacgccgc
     1981 cgccacagag gtggtgaata tcctgaggtc tctgggcgtg gtgttccgcc tgagcaacac
     2041 cgtgctggga ctgacactgc tggcatgggg caatagcatc ggcgacgcct tctccgactt
     2101 caccctggca aggcagggat acccaagaat ggccttcagc gcctgctttg gcggcatcat
     2161 cttcaacatc ctggtgggag tgggactggg atgtctgctg cagatctcta gaagccacac
     2221 agaggtgaag ctggagccag atggcctgct ggtgtgggtg ctggcaggcg ccctgggact
     2281 gagcctggtg tttagcctgg tgtccgtgcc cctgcagtgc ttccagctgt ccagggtgta
     2341 cggcttttgt ctgctgctgt tctatctgaa ttttctggtg gtggccctgc tgaccgagtt
     2401 cggcgtgatc cacctgaagt ccatggaccc agctttcttg tacaaagttg gcattataag
     2461 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2521 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2581 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2641 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2701 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2761 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2821 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2881 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2941 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     3001 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3061 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3121 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3181 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3241 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3301 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3361 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3421 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3481 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3541 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3601 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3781 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4261 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
