LOCUS       sequence-333112-        3888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3888)
  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 3888)
  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-SLC52A2_STOP
FEATURES             Location/Qualifiers
     source          1..3888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1477..1576)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1593..1612)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1594..1612)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1617..1633)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1617..1633)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1735..1754)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1746..2555
                     /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(1813..1832)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2702..3290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3191..3210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3444..3461
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3620..3647)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3739..3825)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3874..3888,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3888,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacac attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccaccatg gcagcaccta ccccagcaag gcccgtgctg acacacctgc
      181 tggtggccct gttcggaatg ggatcctggg cagcagtgaa cggaatctgg gtggagctgc
      241 ctgtggtggt gaaggagctg ccagagggat ggagcctgcc atcctacgtg tctgtgctgg
      301 tggccctggg caatctggga ctgctggtgg tgaccctgtg gcggagactg gcacctggca
      361 aggatgagca ggtgccaatc agggtggtgc aggtgctggg aatggtggga acagccctgc
      421 tggcaagcct gtggcaccac gtggcaccag tggcaggaca gctgcactcc gtggccttcc
      481 tggccctggc ctttgtgctg gccctggcat gctgtgcctc caacgtgacc ttcctgcctt
      541 ttctgtctca cctgccccct aggttcctgc gctccttctt tctgggacag ggactgtctg
      601 ccctgctgcc atgcgtgctg gccctggtgc agggagtggg ccggctggag tgtccaccag
      661 cacccatcaa tggaacccct ggacctccac tggatttcct ggagagattt ccagcctcca
      721 ccttcttttg ggccctgaca gccctgctgg tggcatctgc cgcagcattt cagggactgc
      781 tgctgctgct gccccctcca cccagcgtgc ccacaggcga gctgggctcc ggactgcaag
      841 tgggagcacc aggagcagag gaggaggtgg aggagagctc cccactgcag gagcctccat
      901 cccaggcagc aggaaccaca ccaggacctg acccaaaggc ctaccagctg ctgagcgccc
      961 ggtccgcctg cctgctggga ctgctggccg ccaccaacgc cctgacaaat ggcgtgctgc
     1021 ccgccgtgca gtccttctct tgtctgcctt acggcaggct ggcctatcac ctggcagtgg
     1081 tgctgggaag cgccgcaaac cctctggcat gcttcctggc catgggcgtg ctgtgccgct
     1141 ctctggcagg actgggagga ctgagcctgc tgggcgtgtt ttgcggagga tacctgatgg
     1201 cactggccgt gctgagccca tgtccacctc tggtgggcac ctccgccgga gtggtgctgg
     1261 tggtgctgtc ttgggtgctg tgcctgggcg tgtttagcta tgtgaaggtg gccgcctcta
     1321 gcctgctgca cggaggaggc cggcctgccc tgctggcagc cggcgtggca atccaagtgg
     1381 gatctctgct gggagcagtg gcaatgttcc cacccacatc tatctatcac gtgtttcaca
     1441 gcagaaagga ctgcgccgat ccctgtgaca gctaagaccc agctttcttg tacaaagttg
     1501 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1561 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     1621 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     1681 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1741 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     1801 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1861 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1921 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1981 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2041 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2101 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2161 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2221 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2281 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2341 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     2401 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2461 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2521 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     2581 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     2641 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3361 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3421 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3481 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3541 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     3601 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     3661 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     3721 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     3781 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     3841 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgtt
//
