LOCUS       sequence-262892-        4716 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 4716)
  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 4716)
  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_SLCO4A1
FEATURES             Location/Qualifiers
     source          1..4716
                     /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(2841..2940)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2957..2976)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2958..2976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2981..2997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2981..2997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3099..3118)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3110..3919
                     /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(3177..3196)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4066..4654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4555..4574
                     /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 accatgccac tgcaccagct gggcgacaag cccctgacct ttcctagccc
      721 aaactccgcc atggagaatg gcctggatca caccccacct tctaggagag caagcccagg
      781 aacaccactg tctcctggaa gcctgcggtc cgccgcacac tccccactgg acacctctaa
      841 gcagcctctg tgccagctgt gggcagagaa gcacggagca aggggcacac acgaggtgag
      901 atacgtgtcc gccggacagt ctgtggcatg cggatggtgg gccttcgccc caccatgtct
      961 gcaggtgctg aacaccccca agggcatcct gttcttcctg tgcgcagcag ccttcctgca
     1021 gggaatgaca gtgaacggct ttatcaatac cgtgatcaca agcctggaga ggcgctacga
     1081 cctgcactct tatcagagcg gcctgatcgc cagctcctat gatatcgccg cctgcctgtg
     1141 cctgaccttc gtgagctact ttggaggatc cggacacaag cctaggtggc tgggatgggg
     1201 cgtgctgctg atgggaaccg gatccctggt gttcgcactg ccacacttta cagccggcag
     1261 gtatgaggtg gagctggacg caggcgtgcg cacctgccca gcaaatccag gagccgtgtg
     1321 cgcagattcc acatctggcc tgagccgcta ccagctggtg ttcatgctgg gacagtttct
     1381 gcacggagtg ggagcaacac cactgtacac cctgggcgtg acatatctgg acgagaacgt
     1441 gaagtctagc tgctccccag tgtacatcgc catcttttat accgcagcaa tcctgggacc
     1501 agcagcagga tatctgatcg gaggcgccct gctgaatatc tacaccgaga tgggccggag
     1561 aacagagctg accacagagt ccccactgtg ggtgggagca tggtgggtgg gcttcctggg
     1621 atctggagca gcagccttct ttacagcagt gccaatcctg ggctatccca gacagctgcc
     1681 tggcagccag aggtacgccg tgatgagggc agcagagatg caccagctga aggattcctc
     1741 taggggagag gcaagcaacc cagacttcgg caagaccatc cgcgatctgc ctctgtccat
     1801 ctggctgctg ctgaagaatc ccaccttcat cctgctgtgc ctggcaggag caaccgaggc
     1861 cacactgatc accggcatgt ccaccttctc tcctaagttt ctggagtccc agttcagcct
     1921 gtccgcctct gaggcagcca ccctgtttgg ctatctggtg gtgccagcag gaggaggagg
     1981 cacatttctg ggcggcttct ttgtgaacaa gctgaggctg cgcggctctg ccgtgatcaa
     2041 gttctgcctg ttttgtaccg tggtgagcct gctgggcatc ctggtgttct ctctgcactg
     2101 ccctagcgtg ccaatggcag gagtgaccgc aagctatgga ggatccctgc tgcctgaggg
     2161 acacctgaac ctgacagccc catgcaatgc cgcctgctct tgtcagccag agcactattc
     2221 ccccgtgtgc ggctctgacg gcctgatgta ctttagcctg tgccacgcag gatgtccagc
     2281 agcaaccgag acaaacgtgg acggacagaa ggtgtaccgg gattgctcct gtatccctca
     2341 gaatctgagc tccggattcg gacacgcaac cgccggcaag tgcacctcta catgtcagag
     2401 aaagcccctg ctgctggtgt tcatctttgt ggtcatcttc ttcacattcc tgtctagcat
     2461 ccccgccctg accgccacac tgagatgcgt gagagatcct cagcggagct ttgccctggg
     2521 catccagtgg atcgtggtga gaatcctggg aggaatccca ggacctatcg ccttcggctg
     2581 ggtcatcgac aaggcctgtc tgctgtggca ggatcagtgc ggccagcagg gctcctgtct
     2641 ggtgtaccag aatagcgcca tgtccaggta tatcctgatc atgggcctgc tgtacaaggt
     2701 gctgggcgtg ctgttctttg ccatcgcctg cttcctgtac aagcctctga gcgagtcctc
     2761 tgatggcctg gagacctgtc tgccttccca gagctccgcc ccagactctg ccacagatag
     2821 ccagctgcag tctagcgtgg acccagcttt cttgtacaaa gttggcatta taagaaagca
     2881 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     2941 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3001 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3061 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3121 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3181 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3241 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3301 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3361 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3421 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3481 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3541 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3601 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3661 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3721 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3781 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3841 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3901 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     3961 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     4021 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4081 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4141 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4201 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4261 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4321 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4381 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4441 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4501 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4561 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4621 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4681 gttcctggcc ttttgctggc cttttgctca catgtt
//
