LOCUS       sequence-260402-        4242 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 4242)
  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 4242)
  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_SLC1A6
FEATURES             Location/Qualifiers
     source          1..4242
                     /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(2367..2466)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2483..2502)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2484..2502)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2507..2523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2507..2523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2625..2644)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2636..3445
                     /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(2703..2722)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3592..4180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4081..4100
                     /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 accatgagct cccacggcaa ctccctgttc ctgagggagt ctggacagcg
      721 cctgggaagg gtgggatggc tgcagcggct gcaggagagc ctgcagcaga gagccctgcg
      781 gaccagactg aggctgcaga ccatgacact ggagcacgtg ctgagattcc tgcggagaaa
      841 tgcctttatc ctgctgaccg tgtctgccgt ggtcatcggc gtgagcctgg cctttgccct
      901 gaggccatac cagctgacat atcgccagat caagtacttc tcctttcccg gcgagctgct
      961 gatgaggatg ctgcagatgc tggtgctgcc tctgatcgtg tctagcctgg tgaccggcat
     1021 ggcctctctg gacaacaagg ccacaggcag aatgggcatg agggccgccg tgtactatat
     1081 ggtgaccaca atcatcgccg tgttcatcgg catcctgatg gtgaccatca tccaccctgg
     1141 caagggcagc aaggagggcc tgcacagaga gggcaggatc gagaccatcc caacagccga
     1201 cgccttcatg gatctgatca ggaacatgtt tccccctaat ctggtggagg cctgcttcaa
     1261 gcagtttaag acccagtact ccacacgcgt ggtgacccgg acaatggtga gaaccgagaa
     1321 tggatctgag cctggagcaa gcatgccacc acctttcagc gtggagaacg gcacatcctt
     1381 tctggagaat gtgacccgcg ccctgggcac actgcaggag atgctgtcct tcgaggagac
     1441 cgtgccagtg ccaggatctg ccaacggcat caatgccctg ggcctggtgg tgttctctgt
     1501 ggcctttggc ctggtcatcg gcggcatgaa gcacaagggc cgcgtgctgc gggacttctt
     1561 tgatagcctg aacgaggcca tcatgcggct ggtgggcatc atcatctggt atgccccagt
     1621 gggcatcctg tttctgatcg ccggcaagat cctggagatg gaggatatgg ccgtgctggg
     1681 aggacagctg ggaatgtaca ccctgacagt gatcgtgggc ctgttcctgc acgccggcat
     1741 cgtgctgccc ctgatctatt ttctggtgac ccacagaaat cctttcccct tcatcggagg
     1801 aatgctgcag gccctgatca ccgccatggg cacatcctct agctccgcca ccctgcccat
     1861 cacattcagg tgtctggagg agggactggg agtggacagg aggatcacac gctttgtgct
     1921 gcctgtgggc gccaccgtga acatggatgg aacagccctg tacgaggccc tggcagcaat
     1981 cttcatcgcc caggtgaaca attatgagct gaatctgggc cagatcacca caatctccat
     2041 caccgcaaca gcagcatctg tgggagcagc aggcatccca caggccggcc tggtgaccat
     2101 ggtcatcgtg ctgaccagcg tgggactgcc aaccgaggac atcacactga tcatcgccgt
     2161 ggactggttt ctggatcgcc tgcggaccat gacaaacgtg ctgggcgata gcatcggagc
     2221 agccgtgatc gagcacctgt cccagaggga gctggagctg caggaggcag agctgaccct
     2281 gccaagcctg ggcaagcctt ataagtccct gatggcccag gagaagggag catctagagg
     2341 aaggggaggc aatgagagcg ccatggaccc agctttcttg tacaaagttg gcattataag
     2401 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2461 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2521 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2581 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     2641 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     2701 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     2761 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2821 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2881 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     2941 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3001 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3061 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3121 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3181 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3241 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3301 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3361 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3421 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     3481 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     3541 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3601 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3661 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3721 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3781 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3841 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3901 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3961 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4021 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4081 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4141 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4201 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
