LOCUS       sequence-253297-        4479 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 4479)
  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 4479)
  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_SLCO2A1
FEATURES             Location/Qualifiers
     source          1..4479
                     /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(2604..2703)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2720..2739)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2721..2739)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2744..2760)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2744..2760)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2862..2881)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2873..3682
                     /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(2940..2959)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3829..4417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4318..4337
                     /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 accatgggcc tgctgccaaa gctgggagca tctcagggaa gcgacacctc
      721 tacaagcagg gcaggcagat gcgcacggag cgtgttcggc aacatcaagg tgttcgtgct
      781 gtgccaggga ctgctgcagc tgtgccagct gctgtacagc gcctacttca agagctccct
      841 gaccacaatc gagaagcggt tcggcctgtc tagctcctct agcggcctga tctcctctct
      901 gaacgagatc tccaatgcca tcctgatcat cttcgtgtct tactttggaa gcagggtgca
      961 caggccaaga ctgatcggaa tcggaggact gtttctggca gcaggagcct tcatcctgac
     1021 cctgccccac tttctgagcg agccttacca gtataccctg gcctctacag gcaacaatag
     1081 caggctgcag gcagagctgt gccagaagca ctggcaggat ctgccccctt ccaagtgcca
     1141 ctctaccaca cagaaccccc agaaggagac cagctccatg tggggactga tggtggtggc
     1201 acagctgctg gcaggaatcg gcacagtgcc aatccagccc tttggcatct cctacgtgga
     1261 cgatttctct gagccatcca attctcccct gtatatcagc atcctgtttg ccatctccgt
     1321 gttcggccct gcctttggct acctgctggg ctccgtgatg ctgcagatct tcgtggacta
     1381 tggccgcgtg aacaccgccg ccgtgaatct ggtgcctggc gatccaaggt ggatcggagc
     1441 atggtggctg ggactgctga tctctagcgc cctgctggtg ctgaccagct tccccttctt
     1501 tttctttcct agagccatgc caatcggagc aaagagggca cctgcaacag cagacgaggc
     1561 aaggaagctg gaggaggcaa agtcccgggg ctctctggtg gacttcatca agcggttccc
     1621 ttgtatcttc ctgcggctgc tgatgaactc cctgtttgtg ctggtggtgc tggcccagtg
     1681 caccttctcc tctgtgatcg ccggcctgag cacatttctg aataagttcc tggagaagca
     1741 gtacggcacc tccgccgcct atgccaactt tctgatcgga gcagtgaatc tgcctgcagc
     1801 cgccctggga atgctgttcg gaggaatcct gatgaagaga ttcgtgttta gcctgcaggc
     1861 catcccaagg atcgccacca caatcatcac catctccatg atcctgtgcg tgcctctgtt
     1921 ctttatgggc tgctctaccc caacagtggc cgaggtgtac ccacccagca caagctcctc
     1981 tatccacccc cagtcccctg cctgtcggag agactgcagc tgtccagatt ccatttttca
     2041 ccccgtgtgc ggcgacaacg gcatcgagta cctgtcccca tgccacgcag gatgttctaa
     2101 catcaatatg agctccgcca cctctaagca gctgatctat ctgaattgca gctgcgtgac
     2161 cggaggaagc gcctccgcca agacaggatc ctgccctgtg ccatgtgccc acttcctgct
     2221 gccagccatc tttctgatca gcttcgtgtc cctgatcgcc tgtatctctc acaaccccct
     2281 gtacatgatg gtgctgagag tggtgaatca ggaggagaag agctttgcca tcggcgtgca
     2341 gttcctgctg atgaggctgc tggcatggct gccatcccct gccctgtatg gcctgacaat
     2401 cgaccactct tgcatcagat ggaacagcct gtgcctgggc aggcgcggag catgcgccta
     2461 ctatgacaat gatgccctga gggatcgcta cctgggcctg cagatgggct ataaggccct
     2521 gggcatgctg ctgctgtgct tcatcagctg gagggtgaag aagaacaagg agtataatgt
     2581 gcagaaggcc gccggcctga tcgacccagc tttcttgtac aaagttggca ttataagaaa
     2641 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2701 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2761 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaataatatc
     2821 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2881 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2941 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3001 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3061 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3121 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3181 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3241 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3301 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3361 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3421 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3481 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3541 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3601 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3661 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3721 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3781 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4441 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
