LOCUS       sequence-373230-        3960 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 3960)
  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 3960)
  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-SLC52A3_STOP
FEATURES             Location/Qualifiers
     source          1..3960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1517..1616)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1633..1652)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1634..1652)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1657..1673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1657..1673)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1775..1794)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1786..2595
                     /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(1853..1872)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2742..3330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3231..3250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3484..3501
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3660..3687)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3779..3865)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3914..3936
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3928..3945
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3929..3945
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cttcctgatg
      121 cacctgctgg tgtgcgtgtt tggcatgggc agctgggtga ccatcaacgg cctgtgggtg
      181 gagctgcctc tgctggtcat ggagctgcca gagggctggt acctgccctc ctatctgaca
      241 gtggtcatcc agctggccaa tatcggccca ctgctggtga ccctgctgca ccacttccgg
      301 ccatcctgcc tgtctgaggt gcctatcatc tttaccctgc tgggcgtggg caccgtgaca
      361 tgtatcatct tcgcctttct gtggaacatg acatcttggg tgctggacgg ccaccacagc
      421 atcgccttcc tggtgctgac cttctttctg gccctggtgg attgcaccag ctccgtgaca
      481 ttcctgcctt ttatgagcag actgccaaca tactatctga ccacattctt tgtgggcgag
      541 ggcctgtctg gactgctgcc cgccctggtg gccctggcac agggcagcgg cctgaccaca
      601 tgcgtgaatg tgaccgagat cagcgactcc gtgccctctc ctgtgccaac gagggaaacg
      661 gatatagctc agggagtgcc tcgcgccctg gtgagcgccc tgcctggcat ggaggcccca
      721 ctgtcccacc tggagtctcg gtatctgccc gcccacttta gccctctggt gttctttctg
      781 ctgctgtcca tcatgatggc ctgctgtctg gtggccttct ttgtgctgca gaggcagcca
      841 aggtgctggg aggcatccgt ggaggacctg ctgaacgatc aggtgacact gcactctatc
      901 cggccaagag aggagaatga cctgggacca gcaggaaccg tggattctag ccagggccag
      961 ggctatctgg aggagaaggc agcaccatgc tgtccagcac acctggcctt catctacacc
     1021 ctggtggcct ttgtgaacgc cctgacaaat ggcatgctgc cctccgtgca gacctacagc
     1081 tgtctgtcct atggacctgt ggcataccac ctggcagcca cactgtctat cgtggccaac
     1141 cctctggcct ctctggtgag catgttcctg ccaaataggt ccctgctgtt tctgggcgtg
     1201 ctgtctgtgc tgggcacctg cttcggcggc tataacatgg ccatggccgt gatgagcccc
     1261 tgtcctctgc tgcagggaca ctggggagga gaggtgctga tcgtggcatc ctgggtgctg
     1321 tttagcggct gcctgtccta cgtgaaagtg atgctgggcg tggtgctgcg ggacctgtct
     1381 agaagcgccc tgctgtggtg tggagcagca gtgcagctgg gctccctgct gggcgccctg
     1441 ctgatgttcc cactggtgaa cgtgctgcgc ctgttctcct ctgccgattt ttgcaatctg
     1501 cactgtcccg cctaagaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1561 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1621 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1681 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1741 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1801 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1861 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1921 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1981 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2041 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2101 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2161 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2221 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2281 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2341 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2401 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2461 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2521 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2581 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2641 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2881 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3421 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3481 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3541 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3601 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     3661 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     3721 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     3781 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     3841 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     3901 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
