LOCUS       sequence-479352-        3924 bp    DNA     circular SYN 28-JUL-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 3924)
  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 3924)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 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-UNC93A_STOP
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(76..103)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(195..281)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     330..352
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     344..361
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     345..361
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    377..476
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1743..1752
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1857..1956)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1973..1992)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1974..1992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1997..2013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1997..2013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2115..2134)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2126..2935
                     /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(2193..2212)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3082..3670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3571..3590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3824..3841
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga
       61 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg
      121 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa
      181 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
      241 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
      301 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
      361 tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct gttcgttgca
      421 acacattgat gagcaatgct tttttataat gccaactttg tacaaaaaag caggctccac
      481 catggacagg tccctgcgca acgtgctggt ggtgtctttc ggctttctgc tgctgttcac
      541 cgcatacgga ggactgcaga gcctgcagag ctccctgtat tccgaggagg gactgggagt
      601 gaccgccctg tccacactgt acggcggcat gctgctgtct agcatgttcc tgccacctct
      661 gctgatcgag cggctgggat gcaagggcac catcatcctg agcatgtgcg gctacgtggc
      721 cttttctgtg ggcaatttct ttgccagctg gtataccctg atccctacaa gcatcctgct
      781 gggactggga gcagcaccac tgtggtccgc ccagtgcacc tacctgacca tcacaggcaa
      841 cacacacgcc gagaaggccg gcaagagagg caaggacatg gtgaatcagt atttcggcat
      901 cttctttctg atctttcagt cctctggcgt gtggggcaac ctgatcagct ccctggtgtt
      961 cggccagacc ccatcccagg agacactgcc agaggagcag ctgacctcct gcggagcctc
     1021 tgattgtctg atggccacca caaccacaaa ttctacacag cggccaagcc agcagctggt
     1081 gtacaccctg ctgggcatct atacaggcag cggcgtgctg gccgtgctga tgatcgcagc
     1141 cttcctgcag cccatcaggg acgtgcagag ggagtccgag ggagagaaga agtctgtgcc
     1201 tttctggagc accctgctga gcaccttcaa gctgtaccgg gataagagac tgtgcctgct
     1261 gatcctgctg cccctgtatt ctggcctgca gcagggcttt ctgtctagcg agtacacccg
     1321 gagctatgtg acctgtacac tgggcatcca gttcgtgggc tacgtgatga tctgcttttc
     1381 cgccacagac gccctgtgct ctgtgctgta cggcaaggtg agccagtata ccggcagagc
     1441 cgtgctgtat gtgctgggcg ccgtgacaca cgtgagctgt atgatcgccc tgctgctgtg
     1501 gaggcctaga gcagaccacc tggccgtgtt cttcgtgttc agcggactgt ggggagtggc
     1561 agatgccgtg tggcagaccc agaacaatgc cctgtacggc gtgctgttcg agaagtctaa
     1621 ggaggccgcc tttgccaact ataggctgtg ggaggccctg ggcttcgtga tcgcctttgg
     1681 ctacagcatg tttctgtgcg tgcacgtgaa gctgtatatc ctgctgggcg tgctgtccct
     1741 gaccatggtg gcctacggcc tggtggagtg cgtggagtct aagaacccaa tcaggccaca
     1801 cgcacctgga caagtgaatc aggccgagga tgaggagatc cagacaaaga tgtaagaccc
     1861 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1921 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     1981 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2041 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     2101 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     2161 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     2221 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     2281 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     2341 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     2401 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     2461 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     2521 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     2581 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     2641 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     2701 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     2761 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     2821 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     2881 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     2941 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     3001 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3061 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3841 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3901 atgcagctgg cacgacaggt ttcc
//
