LOCUS       sequence-260206-        4209 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 4209)
  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 4209)
  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_SLC22A12
FEATURES             Location/Qualifiers
     source          1..4209
                     /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(2334..2433)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2450..2469)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2451..2469)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2474..2490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2474..2490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2592..2611)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2603..3412
                     /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(2670..2689)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3559..4147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4048..4067
                     /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 accatggcct tttccgagct gctggacctg gtgggaggac tgggccggtt
      721 ccaggtgctg cagaccatgg ccctgatggt gagcatcatg tggctgtgca cacagtccat
      781 gctggagaac tttagcgccg cagtgccatc ccacagatgt tgggcccctc tgctggataa
      841 ttctaccgcc caggccagca tcctgggatc tctgagccct gaggccctgc tggcaatcag
      901 catcccacct ggacctaacc agaggccaca ccagtgccgg cggttccggc agccacagtg
      961 gcagctgctg gaccccaatg ccaccgccac atcctggtct gaggcagata ccgagccatg
     1021 cgtggacgga tgggtgtacg ataggagcat cttcacctcc acaatcgtgg ccaagtggaa
     1081 cctggtgtgc gactctcacg ccctgaagcc aatggcccag agcatctatc tggccggcat
     1141 cctggtggga gcagcagcat gcggaccagc aagcgatagg tttggcaggc gcctggtgct
     1201 gacctggtcc tacctgcaga tggccgtgat gggaacagca gcagcctttg ccccagcatt
     1261 ccccgtgtac tgtctgttcc gctttctgct ggccttcgcc gtggcaggcg tgatgatgaa
     1321 caccggcaca ctgctgatgg agtggaccgc agcaagggca aggccactgg tcatgaccct
     1381 gaactctctg ggcttcagct ttggccacgg actgaccgca gcagtggcat acggcgtgcg
     1441 ggactggaca ctgctgcagc tggtggtgtc cgtgcccttc ttcctgtgct tcctgtattc
     1501 ctggtggctg gcagagtctg ccaggtggct gctgaccaca ggccgcctgg actggggact
     1561 gcaggagctg tggagagtgg cagccatcaa tggcaagggc gccgtgcagg ataccctgac
     1621 accagaggtg ctgctgagcg ccatgaggga ggagctgtcc atgggacagc caccagcatc
     1681 tctgggcacc ctgctgagaa tgcctggcct gcggttcaga acctgcatca gcaccctgtg
     1741 ctggttcgcc tttggcttca cattctttgg actggccctg gacctgcagg ccctgggatc
     1801 taacatcttt ctgctgcaga tgttcatcgg cgtggtggat atcccagcaa agatgggcgc
     1861 cctgctgctg ctgtcccacc tgggccggag acctaccctg gcagcatctc tgctgctggc
     1921 aggactgtgc atcctggcaa atacactggt gccacacgag atgggcgccc tgaggagcgc
     1981 cctggccgtg ctgggactgg gaggagtggg cgccgccttt acctgtatca caatctacag
     2041 ctccgagctg ttccctaccg tgctgaggat gacagcagtg ggactgggcc agatggcagc
     2101 aaggggagga gcaatcctgg gaccactggt gcgcctgctg ggcgtgcacg gcccatggct
     2161 gcccctgctg gtgtatggca ccgtgcctgt gctgtctgga ctggccgccc tgctgctgcc
     2221 agagacccag agcctgcctc tgccagacac aatccaggat gtgcagaacc aggccgtgaa
     2281 gaaggccacc cacggcacac tgggcaattc cgtgctgaag tctacccagt tcgacccagc
     2341 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2401 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2461 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2521 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2581 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2641 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2701 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2761 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2821 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2881 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2941 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3001 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3061 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3121 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3181 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3241 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3301 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3361 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3421 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3481 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3541 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3601 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3661 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3721 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3781 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3841 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3901 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3961 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4021 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4081 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4141 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4201 tcacatgtt
//
