LOCUS       sequence-261915-        3414 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 3414)
  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 3414)
  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_SLC25A45
FEATURES             Location/Qualifiers
     source          1..3414
                     /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(1539..1638)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1655..1674)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1656..1674)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1679..1695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1679..1695)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1797..1816)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1808..2617
                     /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(1875..1894)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2764..3352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3253..3272
                     /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 accatgccag tggaggagtt cgtggccggc tggatctccg gcgccctggg
      721 actggtgctg ggacacccct ttgacaccgt gaaggtgcgc ctgcagacac agaccacata
      781 ccggggcatc gtggattgca tggtgaagat ctatcgccac gagtctctgc tgggcttctt
      841 taagggcatg tccttcccca tcgcctctat cgccgtggtg aacagcgtgc tgtttggcgt
      901 gtactccaat accctgctgg tgctgaccgc cacatctcac caggagagga gagcacagcc
      961 acctagctac atgcacatct ttttggcagg ttgcacgggt ggatttctgc aggcctattg
     1021 tctggcccct ttcgacctga tcaaggtgag actgcagaac cagacagagc caagggcaca
     1081 gccaggatct ccaccaccta gatatcaggg accagtgcac tgcgcagcaa gcatcttcag
     1141 ggaggaggga cctagaggac tgtttagggg agcatgggca ctgaccctga gggatacccc
     1201 tacagtgggc atctacttca tcacctatga gggcctgtgc cggcagtaca cacctgaggg
     1261 ccagaatcca agctccgcca ccgtgctggt ggcaggagga tttgccggca tcgcatcctg
     1321 ggtggcagca acaccactgg acatgatcaa gtctagaatg cagatggatg gcctgaggcg
     1381 ccgggtgtat cagggcatgc tggactgtat ggtgtctagc atcaggcagg agggcctggg
     1441 cgtgttcttt aggggagtga ccatcaacag cgccagggca ttcccagtga atgcagtgac
     1501 atttctgtcc tacgagtatc tgctgcggtg gtggggcgac ccagctttct tgtacaaagt
     1561 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1621 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1681 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1741 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1801 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1861 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1921 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1981 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2041 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2101 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2161 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2221 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2281 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2341 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2401 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2461 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2521 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2581 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2641 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2701 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
