LOCUS       sequence-259079-        4692 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 4692)
  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 4692)
  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_SLC12A8
FEATURES             Location/Qualifiers
     source          1..4692
                     /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(2817..2916)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2933..2952)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2934..2952)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2957..2973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2957..2973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3075..3094)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3086..3895
                     /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(3153..3172)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4042..4630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4531..4550
                     /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 accatgaccc agatgtccca ggtgcaggag ctgttccacg aggcagcaca
      721 gcaggacgca ctggcacagc cacagccctg gtggaagaca cagctgttca tgtgggagcc
      781 cgtgctgttt ggcacctggg atggcgtgtt cacaagctgc atgatcaaca tcttcggcgt
      841 ggtgctgttt ctgagaaccg gctggctggt gggcaataca ggcgtgctgc tgggcatgtt
      901 cctggtgtcc tttgtgatcc tggtggccct ggtgaccgtg ctgtctggca tcggcgtggg
      961 agagaggagc tccatcggaa gcggaggcgt gtactccatg atctctagcg tgctgggagg
     1021 acagaccgga ggcacaatcg gcctgctgta cgtgtttggc cagtgcgtgg ccggcgccat
     1081 gtatatcaca ggcttcgccg agagcatctc cgacctgctg ggactgggaa acatctgggc
     1141 cgtgcgcgga atctccgtgg ccgtgctgct ggccctgctg ggcatcaatc tggccggcgt
     1201 gaagtggatc atccggctgc agctgctgct gctgtttctg ctggccgtga gcaccctgga
     1261 cttcgtggtg ggctccttta cacacctgga tccagagcac ggcttcatcg gctacagccc
     1321 cgagctgctg cagaacaata ccctgcctga ctattcccca ggcgagtctt tctttaccgt
     1381 gtttggcgtg ttctttccag cagcaacagg cgtgatggca ggcttcaaca tgggaggcga
     1441 tctgagggag ccagcagcat ctatcccact gggaagcctg gcagcagtgg gaatctcctg
     1501 gtttctgtac atcatcttcg tgtttctgct gggagcaatc tgcacaaggg aggccctgag
     1561 atatgacttc ctgatcgccg agaaggtgtc tctgatgggc ttcctgtttc tgctgggcct
     1621 gtacatctcc tctctggcca gctgtatggg aggactgtat ggagcaccaa ggatcctgca
     1681 gtgcatcgca caggagaaag tgatccccgc cctggcatgt ctgggacagg gcaagggccc
     1741 caataagacc cctgtggcag caatctgcct gaccagcctg gtgacaatgg ccttcgtgtt
     1801 tgtgggccag gtgaacgtgc tggcccctat cgtgaccatc aacttcatgc tgacatacgt
     1861 ggccgtggat tactcctatt tctctctgag catgtgctcc tgttctctga cacctgtgcc
     1921 agagcccgtg ctgagagagg gagcagaggg actgcactgt tctgagcacc tgctgctgga
     1981 gaaggcacct agctatggat ccgagggacc agcacagcgg gtgctggagg gcaccctgct
     2041 ggagtttaca aaggacatgg atcagctgct gcagctgacc agaaagctgg agagctccca
     2101 gcctaggcag ggagagggaa acaggacacc agagtcccag aagaggaagt ctaagaaggc
     2161 caccaagcag acactgcagg acagctttct gctggatctg aagtctcccc ctagcttccc
     2221 agtggagatc tccgacaggc tgcctgcagc aagctgggag ggccaggagt cctgctggaa
     2281 caagcagacc agcaagtccg agggaaccca gccagaggga acatacggag agcagctggt
     2341 gcctgagctg tgcaatcagt ctgagtctag cggcgaggac ttctttctga agagcaggct
     2401 gcaggagcag gacgtgtggc ggagatctac cagcttttac acacacatgt gcaatccttg
     2461 ggtgtctctg ctgggagcag tgggaagcct gctgatcatg ttcgtgatcc agtgggtgta
     2521 taccctggtg aacatgggcg tggccgccat cgtgtacttt tatatcggca gagcatcccc
     2581 aggactgcac ctgggatccg cctctaattt ctccttcttt cggtggatga gatctctgct
     2641 gctgccatcc tgtaggtctc tgcgcagccc ccaggagcag atcatcctgg cccctagcct
     2701 ggccaaggtg gatatggaga tgacccagct gacacaggag aacgccgact tcgccaccag
     2761 ggatcgctac caccactcct ctctggtgaa tagggagcag ctgatgcccc actatgaccc
     2821 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2881 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2941 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     3001 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     3061 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     3121 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     3181 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     3241 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3301 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3361 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     3421 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3481 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3541 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3601 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3661 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3721 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3781 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3841 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3901 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3961 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     4021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4681 tgctcacatg tt
//
