LOCUS       sequence-355837-        4137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4137)
  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 4137)
  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-SPNS1_STOP
FEATURES             Location/Qualifiers
     source          1..4137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1694..1793)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1810..1829)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1811..1829)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1834..1850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1834..1850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1952..1971)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1963..2772
                     /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(2030..2049)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2919..3507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3408..3427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3661..3678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3837..3864)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3956..4042)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4091..4113
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4105..4122
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4106..4122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc cggctctgac
      121 accgcccctt tcctgagcca ggccgacgat ccagacgatg gaccagtgcc aggaacccct
      181 ggactgccag gctctacagg caaccccaag agcgaggagc cagaggtgcc tgaccaggag
      241 ggactgcaga ggatcacagg actgtcccct ggccggtctg ccctgatcgt ggccgtgctg
      301 tgctacatca acctgctgaa ttatatggat cgctttaccg tggcaggcgt gctgccagac
      361 atcgagcagt tctttaacat cggcgatagc tcctctggcc tgatccagac agtgttcatc
      421 agctcctaca tggtgctggc ccccgtgttc ggctatctgg gcgatcgcta caatcggaag
      481 tatctgatgt gcggcggcat cgccttctgg agcctggtga ccctgggctc tagcttcatc
      541 ccaggcgagc acttttggct gctgctgctg acaaggggac tggtgggagt gggagaggca
      601 agctactcca ccatcgcacc tacactgatc gccgacctgt ttgtggccga tcagagaagc
      661 aggatgctgt ccatcttcta ctttgccatc ccagtgggaa gcggactggg atatatcgca
      721 ggatccaagg tgaaggacat ggccggcgat tggcactggg cactgagagt gacccctgga
      781 ctgggagtgg tggccgtgct gctgctgttc ctggtggtga gagagccacc taggggagca
      841 gtggagaggc actctgacct gccacccctg aacccaacaa gctggtgggc agatctgcgc
      901 gccctggcaa ggaatccatc cttcgtgctg tcctctctgg gcttcaccgc cgtggccttt
      961 gtgacaggat ccctggccct gtgggcacct gcctttctgc tgcgctctag ggtggtgctg
     1021 ggagagaccc ctccatgcct gccaggcgac tcctgtagct cctctgattc tctgatcttt
     1081 ggcctgatca cctgcctgac aggcgtgctg ggagtgggac tgggagtgga gatcagccgg
     1141 agactgagac actccaaccc aagggcagac ccactggtgt gcgcaaccgg actgctgggc
     1201 tccgccccct tcctgtttct gtctctggca tgtgcaaggg gaagcatcgt ggcaacctac
     1261 atcttcatct ttatcggcga gacactgctg agcatgaatt gggccatcgt ggccgatatc
     1321 ctgctgtatg tggtcatccc aaccaggcgc tccacagccg aggccttcca gatcgtgctg
     1381 tctcacctgc tgggcgacgc aggaagccca tatctgatcg gcctgatctc cgatcgcctg
     1441 cggagaaatt ggcccccttc tttcctgagc gagtttcggg ccctgcagtt cagcctgatg
     1501 ctgtgcgcct ttgtgggcgc cctgggagga gcagccttcc tgggcaccgc catctttatc
     1561 gaggccgaca ggcgccgggc acagctgcac gtgcagggac tgctgcacga ggcaggctcc
     1621 accgacgata gaatcgtggt gccccagaga ggcaggtcca caagggtgcc tgtggcctct
     1681 gtgctgatct aagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1741 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1801 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1861 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1921 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1981 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2041 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2101 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2161 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2221 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2281 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2341 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2401 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2461 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2521 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2581 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2641 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2701 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2761 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2821 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2881 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2941 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3001 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3061 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3121 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3181 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3241 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3301 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3361 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     3421 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3481 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3541 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3601 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3661 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3721 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3781 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3841 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3901 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3961 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4021 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4081 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
