LOCUS       sequence-265752-        4740 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 4740)
  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 4740)
  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_SLC27A3
FEATURES             Location/Qualifiers
     source          1..4740
                     /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(2865..2964)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2981..3000)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2982..3000)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3005..3021)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3005..3021)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3123..3142)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3134..3943
                     /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(3201..3220)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4090..4678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4579..4598
                     /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 accatgggcg tgtgccagag gaccagggca ccttggaagg agaagtctca
      721 gctggagagg gccgccctgg gcttccgcaa gggaggctct ggcatgtttg ccagcggctg
      781 gaaccagaca gtgccaatcg aggaggcagg atccatggcc gccctgctgc tgctgcctct
      841 gctgctgctg ctgccactgc tgctgctgaa gctgcacctg tggccacagc tgaggtggct
      901 gcctgcagat ctggccttcg ccgtgcgggc cctgtgctgt aagagagccc tgagggcaag
      961 agccctggca gcagcagcag ccgacccaga gggccccgag ggcggctgta gcctggcctg
     1021 gcggctggcc gagctggccc agcagagagc cgcccacacc ttcctgatcc acggctcccg
     1081 gagattttct tacagcgagg cagagaggga gtctaatagg gcagcaagag ccttcctgag
     1141 agccctgggc tgggattggg gccccgacgg cggcgatagc ggcgagggct ccgccggcga
     1201 gggcgagagg gccgcccctg gcgccggcga tgccgccgcc ggcagcggcg ccgagtttgc
     1261 cggcggcgac ggagcagcaa ggggaggagg agcagcagcc cctctgtccc caggcgccac
     1321 cgtggccctg ctgctgccag caggacctga gttcctgtgg ctgtggtttg gactggcaaa
     1381 ggcaggactg aggaccgcct tcgtgccaac agccctgagg aggggccctc tgctgcactg
     1441 cctgcggtct tgtggagcaa gagccctggt gctggcacca gagtttctgg agagcctgga
     1501 gccagacctg cccgccctgc gcgccatggg cctgcacctg tgggcagcag gaccaggaac
     1561 ccaccctgca ggcatctccg atctgctggc agaggtgtct gccgaggtgg acggaccagt
     1621 gcctggatac ctgagctccc cacagtctat caccgataca tgcctgtata tcttcacaag
     1681 cggaaccaca ggactgccaa aggcagcaag aatcagccac ctgaagatcc tgcagtgcca
     1741 gggcttttat cagctgtgcg gcgtgcacca ggaggacgtg atctacctgg ccctgcctct
     1801 gtatcacatg tccggctctc tgctgggcat cgtgggctgt atgggcatcg gcgccaccgt
     1861 ggtgctgaag agcaagttct ccgccggcca gttttgggag gattgccagc agcacagggt
     1921 gacagtgttc cagtacatcg gcgagctgtg ccgctatctg gtgaaccagc cacctagcaa
     1981 ggcagagagg ggacacaagg tgcgcctggc agtgggatcc ggactgaggc cagacacctg
     2041 ggagcggttc gtgcggcggt tcggaccact gcaggtgctg gagacctacg gcctgacaga
     2101 gggcaacgtg gccaccatca attatacagg acagagggga gcagtgggaa gagcaagctg
     2161 gctgtacaag cacatcttcc ccttttccct gatccggtat gatgtgacca caggcgagcc
     2221 aatcagagac ccacagggac actgcatggc cacctcccct ggagagccag gactgctggt
     2281 ggcaccagtg tctcagcaga gccccttcct gggatacgca ggaggacctg agctggccca
     2341 gggcaagctg ctgaaggacg tgtttcggcc aggcgacgtg ttcttcaata caggcgacct
     2401 gctggtgtgc gacgatcagg gcttcctgag gtttcacgac cgcaccggcg atacattcag
     2461 atggaagggc gagaacgtgg ccaccacaga ggtggccgag gtgttcgagg ccctggattt
     2521 tctgcaggag gtgaacgtgt acggcgtgac cgtgccagga cacgagggaa gggcaggcat
     2581 ggccgccctg gtgctgcgcc caccccacgc cctggacctg atgcagctgt acacacacgt
     2641 gtccgagaac ctgcctccat atgccaggcc tcgctttctg cggctgcagg agtctctggc
     2701 caccacagag accttcaagc agcagaaggt gagaatggcc aatgagggct ttgacccttc
     2761 cacactgtct gatccactgt acgtgctgga ccaggcagtg ggagcatacc tgccactgac
     2821 cacagcaagg tatagcgccc tgctggccgg caacctgcgc atcgacccag ctttcttgta
     2881 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2941 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     3001 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     3061 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     3121 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     3181 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3241 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3301 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3361 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3421 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3481 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3541 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3601 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3661 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3721 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3781 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3841 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3901 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3961 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     4021 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4201 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
