LOCUS       sequence-262973-        4194 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 4194)
  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 4194)
  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_SVOP
FEATURES             Location/Qualifiers
     source          1..4194
                     /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(2319..2418)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2435..2454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2436..2454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2459..2475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2459..2475)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2577..2596)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2588..3397
                     /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(2655..2674)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3544..4132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4033..4052
                     /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 accatggagg aggacctgtt ccagctgcgc cagctgcccg tggtgaagtt
      721 tcggagaacc ggagagagcg cccggtccga ggacgataca gcaagcggag agcacgaggt
      781 gcagatcgag ggagtgcacg tgggactgga ggcagtggag ctggacgatg gagcagccgt
      841 gcctaaggag ttcgccaacc caaccgacga taccttcatg gtggaggacg cagtggaggc
      901 aatcggattc ggcaagtttc agtggaagct gtctgtgctg accggactgg catggatggc
      961 agatgcaatg gagatgatga tcctgagcat cctggccccc cagctgcact gtgagtggag
     1021 actgcctagc tggcaggtgg ccctgctgac ctccgtggtg ttcgtgggca tgatgagctc
     1081 ctctacactg tggggcaata tcagcgacca gtacggcagg aagaccggcc tgaagatctc
     1141 cgtgctgtgg acactgtact atggcatcct gagcgccttt gccccagtgt actcctggat
     1201 cctggtgctg aggggactgg tgggattcgg aatcggagga gtgccacagt ccgtgaccct
     1261 gtatgccgag tttctgccca tgaaggcccg ggccaagtgt atcctgctga tcgaggtgtt
     1321 ttgggccatc ggcacagtgt tcgaggtggt gctggccgtg tttgtgatgc cctctctggg
     1381 ctggagatgg ctgctgatcc tgagcgccgt gcctctgctg ctgttcgccg tgctgtgctt
     1441 ttggctgcca gagtctgcca gatatgatgt gctgagcggc aaccaggaga aggccatcgc
     1501 caccctgaag aggatcgcca cagagaatgg cgcccccatg cctctgggca agctgatcat
     1561 cagccggcag gaggacagag gcaagatgag ggatctgttc acccctcact ttcgctggac
     1621 cacactgctg ctgtggttca tctggttttc caacgccttc tcttactatg gcctggtgct
     1681 gctgaccaca gagctgtttc aggcaggcga cgtgtgcgga atcagctccc ggaagaaggc
     1741 agtggaggca aagtgctccc tggcctgtga gtacctgtct gaggaggact atatggatct
     1801 gctgtggacc acactgtccg agttcccagg cgtgctggtg accctgtgga tcatcgatag
     1861 gctgggccgc aagaagacaa tggccctgtg cttcgtgatc ttctcttttt gtagcctgct
     1921 gctgtttatc tgcgtgggcc gcaatgtgct gaccctgctg ctgttcatcg cccgggcctt
     1981 tatctccggc ggcttccagg ccgcctacgt gtatacacca gaggtgtacc caaccgcaac
     2041 aagagccctg ggactgggca cctgttctgg aatggcaagg gtgggcgccc tgatcacacc
     2101 cttcatcgcc caggtcatgc tggagtctag cgtgtacctg accctggccg tgtattccgg
     2161 atgctgtctg ctggccgccc tggcatcttg ctttctgcct atcgagacaa agggaagggg
     2221 actgcaggag tcctctcaca gggagtgggg acaggagatg gtgggaagag gaatgcacgg
     2281 agcaggagtg accaggagca actccggctc tcaggaggac ccagctttct tgtacaaagt
     2341 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2401 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2461 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2521 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2581 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2641 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2701 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2761 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2821 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2881 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2941 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3001 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3061 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3121 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3181 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3241 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3301 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3361 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3421 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3481 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
