LOCUS       sequence-347622-        4329 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 4329)
  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 4329)
  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-SLC6A20_STOP
FEATURES             Location/Qualifiers
     source          1..4329
                     /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(1886..1985)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2002..2021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2003..2021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2026..2042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2026..2042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2144..2163)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2155..2964
                     /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(2222..2241)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3111..3699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3600..3619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3853..3870
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4029..4056)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4148..4234)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4283..4305
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4297..4314
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4298..4314
                     /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 ccaccatgga gaaggcaagg
      121 ccactgtggg caaactccct gcagttcgtg tttgcctgca tctcttacgc agtgggactg
      181 ggaaacgtgt ggaggttccc ttacctgtgc cagatgtatg gcggcggctc ctttctggtg
      241 ccctacatca tcatgctgat cgtggagggc atgcctctgc tgtatctgga gctggcagtg
      301 ggacagagga tgagacaggg aagcatcggc gcctggagga ccatctctcc atacctgagc
      361 ggagtgggag tggcatccgt ggtggtgtct ttctttctga gcatgtacta taacgtgatc
      421 aatgcctggg ccttctggta cctgttccac tcttttcagg acccactgcc ttggagcgtg
      481 tgccccctga acggcaatca caccggctat gatgaggagt gtgagaaggc cagctccacc
      541 cagtactttt ggtatcgcaa gacactgaac atcagcccat ccctgcagga gaatggagga
      601 gtgcagtggg agcccgccct gtgcctgctg ctggcctggc tggtggtgta cctgtgcatc
      661 ctgcggggca ccgagagcac aggcaaggtg gtgtatttca ccgcctccct gccttactgc
      721 gtgctgatca tctatctgat cagaggactg accctgcacg gagcaacaaa cggactgatg
      781 tacatgttta ccccaaagat cgagcagctg gccaacccca aggcctggat caatgccgcc
      841 acacagatct tctttagcct gggactggga ttcggatccc tgatcgcctt tgccagctat
      901 aacgagccat ccaacaattg ccagaagcac gccatcatcg tgtccctgat caattctttc
      961 accagcatct ttgcctctat cgtgacattc agcatctacg gcttcaaggc cacctttaac
     1021 tatgagaatt gtctgaagaa ggtgagcctg ctgctgacca acacattcga cctggaggat
     1081 ggctttctga cagcctctaa tctggagcag gtgaagggct acctggccag cgcctaccct
     1141 tccaagtatt ctgagatgtt cccacagatc aagaactgta gcctggagtc cgagctggac
     1201 accgcagtgc agggaacagg actggccttt atcgtgtaca ccgaggccat caagaacatg
     1261 gaggtgtccc agctgtggtc tgtgctgtat ttctttatgc tgctgatgct gggcatcggc
     1321 tccatgctgg gcaatacagc cgccatcctg acccctctga cagatagcaa gatcatctct
     1381 agccacctgc caaaggaggc catctccggc ctggtgtgcc tggtgaattg tgccatcggc
     1441 atggtgttca ccatggaggc cggcaactac tggttcgaca tctttaatga ttatgccgcc
     1501 accctgagcc tgctgctgat cgtgctggtg gagacaatcg ccgtgtgcta cgtgtatggc
     1561 ctgcggagat ttgagtccga cctgaaggca atgacaggaa gggccgtgtc ttggtactgg
     1621 aaagtgatgt gggccggcgt gtcccctctg ctgatcgtgt ctctgttcgt gttttacctg
     1681 tctgattata tcctgaccgg cacactgaag tatcaggcat gggacgcaag ccagggacag
     1741 ctggtgacca aggattaccc agcatatgca ctggccgtga tcggactgct ggtggcatcc
     1801 tctaccatgt gcatccctct ggccgccctg ggcacattcg tgcagaggcg cctgaagaga
     1861 ggcgacgccg atccagtggc ctaagaccca gctttcttgt acaaagttgg cattataaga
     1921 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     1981 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     2041 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     2101 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     2161 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2221 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2281 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2341 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2401 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2461 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2521 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2581 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2641 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2701 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2761 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2821 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2881 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2941 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3001 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3061 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3241 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3721 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3781 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3841 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     3901 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3961 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     4021 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     4081 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     4141 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     4201 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     4261 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     4321 ctcgggccc
//
