LOCUS       sequence-401521-        4107 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 4107)
  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 4107)
  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-MFSD8_STOP
FEATURES             Location/Qualifiers
     source          1..4107
                     /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(1664..1763)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1780..1799)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1781..1799)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1804..1820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1804..1820)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1922..1941)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1933..2742
                     /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(2000..2019)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2889..3477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3378..3397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3631..3648
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3807..3834)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3926..4012)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4061..4083
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4075..4092
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4076..4092
                     /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 cggcctgagg
      121 aacgagtccg agcaggagcc tctgctgggc gacacaccag gctctcgcga gtgggatatc
      181 ctggagaccg aggagcacta caagtctcgg tggagaagca tccggatcct gtatctgacc
      241 atgttcctga gctccgtggg ctttagcgtg gtcatgatgt ccatctggcc atacctgcag
      301 aagatcgacc ccaccgccga tacaagcttc ctgggctggg tcatcgcctc ctactctctg
      361 ggccagatgg tggccagccc aatctttggc ctgtggtcca attataggcc acgcaaggag
      421 cccctgatcg tgtccatcct gatctctgtg gccgccaact gcctgtacgc ctatctgcac
      481 atccccgcct ctcacaataa gtactatatg ctggtggcaa gaggactgct gggaatcgga
      541 gcaggcaacg tggcagtggt gcggtcttac acagcaggag ccaccagcct gcaggagaga
      601 acatctagca tggccaatat ctccatgtgc caggccctgg gcttcatcct gggacccgtg
      661 ttccagacct gtttcacatt tctgggcgag aagggcgtga cctgggacgt gatcaagctg
      721 cagatcaaca tgtataccac accagtgctg ctgtccgcct tcctgggcat cctgaatatc
      781 atcctgatcc tggccatcct gcgggagcac agagtggacg attccggcag gcagtgtaag
      841 tctatcaact ttgaggaggc ctctacagac gaggcccagg tgccccaggg caacatcgat
      901 caggtggccg tggtggccat caatgtgctg ttctttgtga cactgttcat ctttgccctg
      961 ttcgagacca tcatcacacc tctgaccatg gacatgtacg cctggaccca ggagcaggcc
     1021 gtgctgtata atggcatcat cctggccgcc ctgggagtgg aggcagtggt catcttcctg
     1081 ggcgtgaagc tgctgagcaa gaagatcgga gagagggcaa tcctgctggg aggactgatc
     1141 gtggtgtggg tgggcttctt tatcctgctg ccctggggca accagttccc taagatccag
     1201 tgggaggatc tgcacaacaa ttccatccct aataccacat ttggcgagat catcatcggc
     1261 ctgtggaaga gccctatgga ggacgataac gagaggccaa ccggatgctc catcgagcag
     1321 gcctggtgtc tgtacacacc agtgatccac ctggcccagt tcctgaccag cgccgtgctg
     1381 atcggactgg gatatcccgt gtgcaatctg atgtcttaca cactgtatag caagatcctg
     1441 ggccccaagc ctcagggcgt gtacatggga tggctgaccg caagcggatc cggagcccgg
     1501 atcctgggcc ccatgttcat ctctcaggtg tatgcccact ggggccctag atgggccttt
     1561 agcctggtgt gcggcatcat cgtgctgacc atcacactgc tgggcgtggt gtacaagaga
     1621 ctgatcgccc tgtccgtgag gtacggccgc atccaggagt aagacccagc tttcttgtac
     1681 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1741 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1801 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1861 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     1921 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     1981 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2041 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2101 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2161 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2221 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2281 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2341 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2401 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2461 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2521 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2581 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2641 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2701 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     2761 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     2821 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2881 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2941 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3001 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3061 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3121 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3181 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3241 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3301 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3361 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3421 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3481 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3541 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3601 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3661 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3721 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
     3781 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     3841 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     3901 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     3961 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     4021 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     4081 acgacggcca gtcttaagct cgggccc
//
