LOCUS       sequence-260985-        4392 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 4392)
  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 4392)
  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_SLC6A12
FEATURES             Location/Qualifiers
     source          1..4392
                     /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(2517..2616)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2633..2652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2634..2652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2657..2673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2657..2673)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2775..2794)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2786..3595
                     /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(2853..2872)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3742..4330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4231..4250
                     /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 gcaaggtggc agtgcaggag tgtggaccac ctgccgtgtc
      721 ctgggtgcca gaggagggag agaagctgga ccaggaggac gaggatcagg tgaaggatcg
      781 cggccagtgg accaacaaga tggagtttgt gctgtctgtg gccggcgaga tcatcggcct
      841 gggcaacgtg tggcggttcc cctacctgtg ctataagaat ggcggcggcg ccttctttat
      901 cccctacttt atcttctttt tcgtgtgcgg catccccgtg ttcttcctgg aggtggccct
      961 gggccagtat acctcccagg gctctgtgac agcctggaga aagatctgcc cactgttcca
     1021 gggaatcgga ctggcctccg tggtcatcga gtcttacctg aacgtgtact atatcatcat
     1081 cctggcctgg gccctgtttt atctgttcag ctcctttacc agcgagctgc cttggaccac
     1141 atgtaacaat ttctggaaca ccgagcactg cacagacttt ctgaatcaca gcggcgccgg
     1201 caccgtgaca cctttcgaga acttcacctc cccagtgatg gagttctggg agcggagagt
     1261 gctgggcatc acatccggca tccacgatct gggctctctg agatgggagc tggccctgtg
     1321 cctgctgctg gcctgggtca tctgctactt ttgtatctgg aagggcgtga agtccaccgg
     1381 caaggtggtg tacttcaccg ccacatttcc atatctgatg ctggtcatcc tgctgatcag
     1441 aggagtgaca ctgccaggag catatcaggg aatcatctac tatctgaagc cagatctgtt
     1501 caggctgaag gacccccaag tgtggatgga tgccggcacc cagatctttt tctcttttgc
     1561 catctgccag ggctgtctga cagccctggg cagctacaac aagtatcaca acaattgcta
     1621 caaggactgt atcgccctgt gcttcctgaa tagcgccacc tcctttgtgg ccggcttcgt
     1681 ggtgttttct atcctgggct tcatgagcca ggagcagggc gtgcctatca gcgaggtggc
     1741 agagtccgga ccaggactgg ccttcatcgc ctttcccaag gccgtgacca tgatgcctct
     1801 gtctcagctg tggagctgtc tgtttttcat catgctgatc ttcctgggcc tggacagcca
     1861 gttcgtgtgc gtggagtgtc tggtgacagc ctccatcgat atgttcccta ggcagctgag
     1921 gaagtctggc aggcgcgagc tgctgatcct gaccatcgcc gtgatgtgct acctgatcgg
     1981 cctgtttctg gtgacagagg gcggcatgta tatcttccag ctgtttgatt actatgcctc
     2041 tagcggcatc tgtctgctgt tcctgagcct gtttgaggtg gtgtgcatct cctgggtgta
     2101 cggcgccgac cgcttctatg ataacatcga ggacatgatc ggctaccggc cttggccact
     2161 ggtgaagatc agctggctgt ttctgacccc tggcctgtgc ctggccacat ttctgttctc
     2221 tctgagcaag tacaccccac tgaagtataa caacgtgtac gtgtacccac cctggggcta
     2281 ctccatcggc tggttcctgg ccctgtcctc tatggtgtgc gtgcccctgt ttgtggtcat
     2341 caccctgctg aagacaagag gcccattccg gaagagactg aggcagctga tcacacccga
     2401 cagctccctg ccccagccta agcagcaccc ttgcctggat ggctctgccg gcagaaattt
     2461 cggcccaagc ccaaccaggg agggactgat cgcaggagag aaggagacac acctggaccc
     2521 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     2581 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2641 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2701 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2761 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2821 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2881 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2941 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     3001 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     3061 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     3121 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     3181 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     3241 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     3301 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     3361 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     3421 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     3481 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     3541 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     3601 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3661 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3721 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3781 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3841 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3901 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3961 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4021 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4081 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4141 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4201 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4261 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4321 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4381 tgctcacatg tt
//
