LOCUS       sequence-349393-        3657 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 3657)
  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 3657)
  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_STOP
FEATURES             Location/Qualifiers
     source          1..3657
                     /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(1214..1313)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1330..1349)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1331..1349)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1354..1370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1354..1370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1472..1491)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1483..2292
                     /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(1550..1569)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2439..3027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2928..2947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3181..3198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3357..3384)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3476..3562)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3611..3633
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3625..3642
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3626..3642
                     /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 agcagtggca
      121 ccagcaggac caggcgacag cgcctccgcc gccctggatg agctgtctct gaactttacc
      181 tacggagcac caggagcagg aaatggatct ctgagcggcg actggtatcg gagaaaccag
      241 atccacctgt tcggcgtgct gctggccatc ctgggcaacc tggtcatctc catctctctg
      301 aatatccaga agtacagcca cctgcagctg gcacagcagg agcacccaag gccttatttc
      361 aagtccgtgc tgtggtgggg aggcgtgctg ctgatggcag tgggagagac aggcaacttt
      421 gcagcatacg gattcgcacc aatcaccctg atcgcacctc tgggatgcgt gagcgtgaca
      481 ggatccgcca tcatctctgt gacctttctg aaggacaatc tgagggccag cgatctgctg
      541 ggcaccacac tggcctttgc cggcacctac ctgctggtga acttcgcccc taatatcaca
      601 caggccatct ccgcccgcac cgtgcagtac tatctggtgg gctggcagtt cctgatctat
      661 gtgatcctgg agatcctgat cttttgtatc ctgctgtact tctataagcg gaagggcatg
      721 aagcacatgg tcatcctgct gacactggtg gccatcctgg ccagcctgac cgtgatctcc
      781 gtgaaggccg tgtctggcat gatcacattt tccgtgatgg ataagatgca gctgacctac
      841 ccaatctttt atatcatgtt catcatcatg atcgcctctt gcgtgtttca ggtgaagttc
      901 ctgaaccagg ccacaaagct gtacaatacc acaaccgtgg tgccagtgaa tcacatcttc
      961 tttaccatct ccgccatcat cgccggcatc atcttttacc aggagttcct gggcgccccc
     1021 tttctgacag tgttcatcta tctgttcggc tgttttctgt ctttcctggg cgtgtttctg
     1081 gtgaccagga accgcgagaa ggagcacctg cagcagagct atatcgactt cggcaatatc
     1141 cctgatacaa ccccagagcg gaaggcctgg agagagacaa acgtgggcca gaatacaacc
     1201 agattcacct gagacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     1261 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     1321 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     1381 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1441 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1501 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1561 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     1621 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1681 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1741 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     1801 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1861 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     1921 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     1981 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2041 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2101 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     2161 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     2221 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     2281 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     2341 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     2401 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2461 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2521 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2581 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2641 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2701 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2761 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2821 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2881 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2941 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     3001 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     3061 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     3121 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3181 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3241 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     3301 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     3361 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     3421 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     3481 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     3541 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     3601 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccc
//
