LOCUS       sequence-260121-        3954 bp    DNA     circular SYN 15-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 3954)
  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 3954)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_SLC25A23
FEATURES             Location/Qualifiers
     source          1..3954
                     /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(2079..2178)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2195..2214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2196..2214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2219..2235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2219..2235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2337..2356)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2348..3157
                     /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(2415..2434)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3304..3892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3793..3812
                     /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 accatgaggg gatctccagg cgatgcagag cggagacagc ggtggggcag
      721 actgttcgag gagctggata gcaacaagga cggaagggtg gatgtgcacg agctgaggca
      781 gggactggca aggctgggag gaggcaatcc agatcctgga gcacagcagg gaatcagctc
      841 cgagggcgac gcagatcctg acggaggact ggacctggag gagttcagca ggtacctgca
      901 ggagagggag cagcgcctgc tgctgatgtt tcactccctg gacagaaacc aggatggcca
      961 catcgacgtg tccgagatcc agcagtcttt tagggccctg ggcatctcca tctctctgga
     1021 gcaggccgag aagatcctgc acagcatgga tagagacggc accatgacaa tcgattggca
     1081 ggagtggagg gaccacttcc tgctgcactc cctggagaat gtggaggatg tgctgtattt
     1141 ttggaagcac tctaccgtgc tggatatcgg cgagtgcctg acagtgcctg acgagttctc
     1201 caagcaggag aagctgaccg gcatgtggtg gaagcagctg gtggcaggag cagtggcagg
     1261 agccgtgtcc agaaccggca cagccccact ggacaggctg aaggtgttta tgcaggtgca
     1321 cgcctctaag acaaaccgcc tgaatatcct gggaggactg cggtctatgg tgctggaggg
     1381 aggaatccgg agcctgtgga gaggcaacgg catcaatgtg ctgaagatcg ccccagagag
     1441 cgccatcaag ttcatggcct acgagcagat caagagggca atcctgggac agcaggagac
     1501 cctgcacgtg caggagcggt ttgtggcagg atccctggca ggagcaaccg cacagacaat
     1561 catctaccca atggaggtgc tgaagacccg gctgacactg aggcgcacag gccagtataa
     1621 gggcctgctg gattgtgcaa ggagaatcct ggagagggag ggaccaaggg ccttctacag
     1681 gggctatctg ccaaacgtgc tgggcatcat cccctacgcc ggcatcgacc tggccgtgta
     1741 tgagaccctg aagaattggt ggctgcagca gtactctcac gatagcgccg accctggaat
     1801 cctggtgctg ctggcatgcg gaaccatctc tagcacatgt ggccagatcg cctcttatcc
     1861 cctggccctg gtgaggacac gcatgcaggc acaggcaagc atcgagggag gacctcagct
     1921 gtccatgctg ggcctgctga gacacatcct gtcccaggag ggcatgagag gcctgtatag
     1981 gggcatcgcc cctaacttta tgaaagtgat cccagccgtg agcatctcct acgtggtgta
     2041 tgagaatatg aagcaggccc tgggcgtgac cagcagggac ccagctttct tgtacaaagt
     2101 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2161 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2221 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2281 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2341 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2401 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2461 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2521 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2581 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2641 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2701 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2761 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2821 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2881 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2941 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3001 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3061 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3121 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3181 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3241 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3901 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
