LOCUS       sequence-261961-        4434 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 4434)
  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 4434)
  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_SLC6A18
FEATURES             Location/Qualifiers
     source          1..4434
                     /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(2559..2658)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2675..2694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2676..2694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2699..2715)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2699..2715)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2817..2836)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2828..3637
                     /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(2895..2914)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3784..4372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4273..4292
                     /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 accatggcac acgcaccaga gccagaccct gcagcatgcg acctgggcga
      721 tgagagacca aagtgggata acaaggccca gtacctgctg agctgcaccg gattcgcagt
      781 gggactgggc aatatctgga ggtttcctta cctgtgccag acatatggcg gcggcgcctt
      841 cctgatcccc tatgtgatcg ccctggtgtt cgagggcatc cctatctttc acgtggagct
      901 ggcaatcgga cagaggctga ggaagggatc cgtgggcgtg tggaccgcaa tctctcctta
      961 cctgagcgga gtgggactgg gatgcgtgac cctgtccttc ctgatctctc tgtactataa
     1021 cacaatcgtg gcctgggtgc tgtggtatct gctgaattcc tttcagcacc cactgccatg
     1081 gagctcctgc ccacctgacc tgaaccgcac aggcttcgtg gaggagtgtc agggctctag
     1141 cgccgtgtct tacttttggt atcggcagac cctgaacatc acagccgaca tcaatgattc
     1201 cggctctatc cagtggtggc tgctgatctg cctggcagca agctgggccg tggtgtacat
     1261 gtgcgtgatc cgcggcatcg agaccacagg caaagtgatc tacttcaccg ccctgtttcc
     1321 atatctggtg ctgacaatct tcctgatcag gggactgacc ctgccaggag ccacaaaggg
     1381 cctgatctac ctgtttaccc caaacatgca catcctgcag aatcccagag tgtggctgga
     1441 tgccgccaca cagatcttct ttagcctgtc cctggccttc ggcggacaca tcgcctttgc
     1501 cagctataac tccccccgca atgactgcca gaaggatgcc gtggtcatcg ccctggtgaa
     1561 ccggatgacc agcctgtacg cctccatcgc cgtgttctct gtgctgggct ttaaggccac
     1621 aaatgactat gagcactgtc tggatcggaa catcctgagc ctgatcaatg acttcgattt
     1681 tcctgagcag tctatcagca gagacgatta cccagccgtg ctgatgcacc tgaatgcaac
     1741 ctggccaaag agggtggcac agctgcctct gaaggcatgc ctgctggagg acttcctgga
     1801 taagtccgcc tctggaccag gactggcctt cgtggtgttt accgagacag acctgcacat
     1861 gccaggagca cccgtgtggg ccatgctgtt ctttggcatg ctgttcaccc tgggcctgag
     1921 caccatgttt ggcacagtgg aggccgtgat cacaccactg ctggatgtgg gcgtgctgcc
     1981 aagatgggtg ccaaaggagg ccctgaccgg actggtgtgc ctggtgtgct tcctgtctgc
     2041 cacctgcttt acactgcaga gcggcaacta ctggctggag atcttcgaca actttgccgc
     2101 ctcccctaat ctgctgatgc tggccttcct ggaggtggtg ggcgtggtgt acgtgtatgg
     2161 catgaagagg ttttgtgacg atatcgcatg gatgaccggc cggagacctt ccccatactg
     2221 gaggctgacc tggcgcgtgg tgtctcctct gctgctgaca atcttcgtgg cctatatcat
     2281 cctgctgttt tggaagccac tgcgctacaa ggcctggaat cccaagtatg agctgttccc
     2341 tagcagacag gagaagctgt atccaggatg ggcaagggca gcatgcgtgc tgctgtccct
     2401 gctgcctgtg ctgtgggtgc cagtggccgc cctggcacag ctgctgacca ggcgccggag
     2461 aacatggagg gacagagatg caaggccaga caccgatatg agacccgaca ccgatacaag
     2521 gcctgacacc gatatgcgcc cagacacaga tatgcgggac ccagctttct tgtacaaagt
     2581 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2641 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2701 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2761 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2821 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2881 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2941 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     3001 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     3061 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     3121 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     3181 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     3241 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     3301 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     3361 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     3421 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     3481 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     3541 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     3601 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     3661 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3721 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3781 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3841 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3901 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3961 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4021 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4081 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4141 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4201 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4261 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4321 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4381 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
