LOCUS       sequence-262999-        3654 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 3654)
  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 3654)
  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-NIPAL2
FEATURES             Location/Qualifiers
     source          1..3654
                     /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(1779..1878)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1895..1914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1896..1914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1919..1935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1919..1935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2037..2056)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2048..2857
                     /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(2115..2134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3004..3592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3493..3512
                     /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 accatggcag cagtggcacc agcaggacca ggcgacagcg cctccgccgc
      721 cctggatgag ctgtctctga actttaccta cggagcacca ggagcaggaa atggatctct
      781 gagcggcgac tggtatcgga gaaaccagat ccacctgttc ggcgtgctgc tggccatcct
      841 gggcaacctg gtcatctcca tctctctgaa tatccagaag tacagccacc tgcagctggc
      901 acagcaggag cacccaaggc cttatttcaa gtccgtgctg tggtggggag gcgtgctgct
      961 gatggcagtg ggagagacag gcaactttgc agcatacgga ttcgcaccaa tcaccctgat
     1021 cgcacctctg ggatgcgtga gcgtgacagg atccgccatc atctctgtga cctttctgaa
     1081 ggacaatctg agggccagcg atctgctggg caccacactg gcctttgccg gcacctacct
     1141 gctggtgaac ttcgccccta atatcacaca ggccatctcc gcccgcaccg tgcagtacta
     1201 tctggtgggc tggcagttcc tgatctatgt gatcctggag atcctgatct tttgtatcct
     1261 gctgtacttc tataagcgga agggcatgaa gcacatggtc atcctgctga cactggtggc
     1321 catcctggcc agcctgaccg tgatctccgt gaaggccgtg tctggcatga tcacattttc
     1381 cgtgatggat aagatgcagc tgacctaccc aatcttttat atcatgttca tcatcatgat
     1441 cgcctcttgc gtgtttcagg tgaagttcct gaaccaggcc acaaagctgt acaataccac
     1501 aaccgtggtg ccagtgaatc acatcttctt taccatctcc gccatcatcg ccggcatcat
     1561 cttttaccag gagttcctgg gcgccccctt tctgacagtg ttcatctatc tgttcggctg
     1621 ttttctgtct ttcctgggcg tgtttctggt gaccaggaac cgcgagaagg agcacctgca
     1681 gcagagctat atcgacttcg gcaatatccc tgatacaacc ccagagcgga aggcctggag
     1741 agagacaaac gtgggccaga atacaaccag attcaccgac ccagctttct tgtacaaagt
     1801 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1861 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1921 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1981 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2041 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2101 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2161 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2221 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2281 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2341 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2401 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2461 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2521 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2581 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2641 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2701 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2761 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2821 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2881 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2941 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3001 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3061 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
