LOCUS       sequence-262989-        4026 bp    DNA     circular SYN 14-APR-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 4026)
  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 4026)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ANKH
FEATURES             Location/Qualifiers
     source          1..4026
                     /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(2151..2250)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2267..2286)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2268..2286)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2291..2307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2291..2307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2409..2428)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2420..3229
                     /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(2487..2506)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3376..3964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3865..3884
                     /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 accatggtga agttccctgc cctgacccac tactggccac tgatccggtt
      721 tctggtgccc ctgggcatca caaacatcgc catcgacttc ggcgagcagg ccctgaatag
      781 aggcatcgcc gccgtgaagg aggatgccgt ggagatgctg gcctcctacg gcctggccta
      841 ttctctgatg aagttcttta ccggccccat gagcgacttc aagaacgtgg gcctggtgtt
      901 tgtgaattcc aagagggatc gcaccaaggc cgtgctgtgc atggtggtgg caggagcaat
      961 cgcagccgtg ttccacacac tgatcgccta cagcgacctg ggctactata tcatcaacaa
     1021 gctgcaccac gtggatgagt ctgtgggcag caagacccgg agagcctttc tgtacctggc
     1081 cgccttccct tttatggacg ccatggcctg gacacacgcc ggcatcctgc tgaagcacaa
     1141 gtattccttc ctggtgggct gcgcctccat ctctgatgtg atcgcccagg tggtgtttgt
     1201 ggccatcctg ctgcactctc acctggagtg tagggagcct ctgctgatcc caatcctgag
     1261 cctgtatatg ggcgccctgg tgcgctgcac caccctgtgc ctgggctact ataagaacat
     1321 ccacgacatc atccccgata ggagcggacc tgagctggga ggcgacgcaa ccatcaggaa
     1381 gatgctgtcc ttctggtggc cactggccct gatcctggca acacagcgga tctctagacc
     1441 tatcgtgaat ctgtttgtga gcagggatct gggaggaagc tccgccgcaa ccgaggcagt
     1501 ggccatcctg accgccacat acccagtggg ccacatgccc tatggctggc tgaccgagat
     1561 ccgcgccgtg taccctgcct tcgacaagaa caatccaagc aacaagctgg tgagcacctc
     1621 caataccgtg acagccgccc acatcaagaa gttcaccttc gtgtgcatgg ccctgtccct
     1681 gaccctgtgc ttcgtgatgt tttggacacc taacgtgagc gagaagatcc tgatcgacat
     1741 catcggcgtg gatttcgcct ttgccgagct gtgcgtggtg ccactgcgga tcttctcttt
     1801 ctttccagtg cccgtgaccg tgagagccca cctgacaggc tggctgatga ccctgaagaa
     1861 gacatttgtg ctggccccat ctagcgtgct gcggatcatc gtgctgatcg cctccctggt
     1921 ggtgctgcca tatctgggag tgcacggagc caccctggga gtgggctccc tgctggccgg
     1981 ctttgtgggc gagtctacaa tggtggccat cgccgcctgt tacgtgtatc ggaagcagaa
     2041 gaagaagatg gagaacgaga gcgccaccga gggagaggac tccgccatga cagatatgcc
     2101 ccctaccgag gaggtgacag atatcgtgga gatgagggag gagaatgagg acccagcttt
     2161 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2221 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2281 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2341 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2401 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2461 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2521 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2581 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2641 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2701 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2761 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2821 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2881 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2941 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3001 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3061 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3121 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3181 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3241 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3301 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3361 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3421 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3481 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3541 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3601 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3661 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3721 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3781 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3841 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3901 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3961 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4021 catgtt
//
