LOCUS       sequence-349394-        3771 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 3771)
  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 3771)
  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-NIPAL3_STOP
FEATURES             Location/Qualifiers
     source          1..3771
                     /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(1328..1427)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1444..1463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1445..1463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1468..1484)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1468..1484)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1586..1605)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1597..2406
                     /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(1664..1683)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2553..3141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3042..3061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3295..3312
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3471..3498)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3590..3676)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3725..3747
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3739..3756
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3740..3756
                     /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 ccaccatgga cggaagccac
      121 tccgccgccc tgaagctgca gcagctgcca cctaccagct cctctagcgc cgtgtccgag
      181 gcctctttca gctacaagga gaacctgatc ggcgccctgc tggccatctt tggccacctg
      241 gtggtgtcca tcgccctgaa tctgcagaag tactgccaca tccggctggc cggctctaag
      301 gacccaagag cctatttcaa gaccaagaca tggtggctgg gcctgtttct gatgctgctg
      361 ggagagctgg gcgtgttcgc aagctatgcc tttgcccccc tgtccctgat cgtgcctctg
      421 agcgccgtgt ccgtgatcgc ctctgccatc atcggcatca tcttcatcaa ggagaagtgg
      481 aagcccaagg atttcctgcg gagatacgtg ctgagctttg tgggatgcgg actggcagtg
      541 gtgggaacct atctgctggt gacattcgcc cctaactctc acgagaagat gaccggcgag
      601 aatgtgacac ggcacctggt gagctggcca tttctgctgt acatgctggt ggagatcatc
      661 ctgttctgtc tgctgctgta cttttataag gagaagaacg ctaacaatat cgtggtcatc
      721 ctgctgctgg tggccctgct gggcagcatg accgtggtga cagtgaaggc cgtggccggc
      781 atgctggtgc tgtccatcca gggcaatctg cagctggact accctatctt ctatgtgatg
      841 ttcgtgtgca tggtggcaac cgccgtgtac caggcagcct tcctgtctca ggccagccag
      901 atgtacgatt cctctctgat cgcctccgtg ggctatatcc tgtctaccac aatcgccatc
      961 acagccggcg ccatcttcta tctggacttt atcggcgagg atgtgctgca catctgcatg
     1021 ttcgccctgg gctgtctgat cgccttcctg ggcgtgtttc tgatcaccag gaaccgcaag
     1081 aagccaatcc cctttgagcc atacatctcc atggacgcca tgcccggcat gcagaatatg
     1141 cacgataagg gcatgacagt gcagcccgag ctgaaggcct ccttttctta cggcgccctg
     1201 gagaacaatg ataacatcag cgagatctat gcccctgcca ccctgccagt gatgcaggag
     1261 gagcacggaa gcaggtccgc ctctggagtg ccttatcgcg tgctggagca cacaaagaag
     1321 gagtgagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     1381 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat
     1441 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     1501 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1561 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1621 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1681 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1741 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1801 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1861 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1921 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1981 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     2041 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     2101 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     2161 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     2221 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     2281 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     2341 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     2401 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     2461 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     2521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3241 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3301 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3361 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3421 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     3481 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     3541 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     3601 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     3661 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     3721 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
