LOCUS       sequence-253955-        4029 bp    DNA     circular SYN 15-DEC-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 4029)
  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 4029)
  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-ANKH
FEATURES             Location/Qualifiers
     source          1..4029
                     /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(2154..2253)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2270..2289)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2271..2289)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2294..2310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2294..2310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2412..2431)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2423..3232
                     /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(2490..2509)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3379..3967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3868..3887
                     /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 gagaatgagt gagacccagc
     2161 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2221 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2281 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2341 atgttacatt gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat
     2401 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2461 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2521 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2581 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2641 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     2701 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     2761 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2821 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2881 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2941 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3001 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3061 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3121 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3181 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3241 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3301 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3961 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4021 tcacatgtt
//
