LOCUS       sequence-367478-        4770 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 4770)
  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 4770)
  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-LETM1_STOP
FEATURES             Location/Qualifiers
     source          1..4770
                     /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(2327..2426)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2443..2462)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2444..2462)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2467..2483)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2467..2483)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2585..2604)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2596..3405
                     /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(2663..2682)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3552..4140
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4041..4060
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4294..4311
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4470..4497)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4589..4675)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4724..4746
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4738..4755
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4739..4755
                     /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 aagcatcctg
      121 ctgcggtcct gcaggggaag agcaccagca agactgccac ctccaccaag gtacaccgtg
      181 ccaagaggaa gcccaggcga cccagcccac ctgtcttgcg ccagcacact gggcctgagg
      241 aactgtctga atgtgccctt cggctgctgt acccctatcc acccagtgta tacaagctcc
      301 aggggcgatc acctgggatg ctgggcactg aggccagagt gtctgaggat cgtgtcccgc
      361 gccccttgga cctccacatc tgtgggcttt gtggcagtgg gaccacagtg tctgcccgtg
      421 cgcggatggc actctagcag gccagtgaga gacgattctg tggtggagaa gtccctgaag
      481 tctctgaagg acaagaacaa gaagctggag gagggaggac ccgtgtacag ccctccagcc
      541 gaggtggtgg tgaagaagtc cctgggacag cgggtgctgg acgagctgaa gcactactat
      601 cacggcttca gactgctgtg gatcgatacc aagatcgccg cccggatgct gtggagaatc
      661 ctgaatggcc acagcctgac acggagagag aggcgccagt tcctgaggat ctgcgccgat
      721 ctgtttcgcc tggtgccttt cctggtgttt gtggtggtgc ctttcatgga gtttctgctg
      781 ccagtggccg tgaagctgtt ccctaacatg ctgccatcca cctttgagac acagtctctg
      841 aaggaggagc ggctgaagaa ggagctgaga gtgaagctgg agctggccaa gttcctgcag
      901 gacaccatcg aggagatggc cctgaagaat aaggccgcca agggcagcgc caccaaggat
      961 ttttccgtgt tctttcagaa gatcagggag acaggagaga ggccatccaa cgaggagatc
     1021 atgaggttct ctaagctgtt tgaggacgag ctgaccctgg ataatctgac acgccctcag
     1081 ctggtggccc tgtgcaagct gctggagctg cagtccatcg gcaccaacaa tttcctgagg
     1141 tttcagctga caatgaggct gcgctccatc aaggccgacg ataagctgat cgccgaggag
     1201 ggcgtggact ctctgaacgt gaaggagctg caggcagcat gtagggcaag aggaatgcgg
     1261 gccctgggcg tgaccgagga caggctgagg ggacagctga agcagtggct ggatctgcac
     1321 ctgcaccagg agatccctac ctctctgctg atcctgagca gagccatgta tctgcccgac
     1381 acactgtctc ctgccgatca gctgaagagc accctgcaga cactgccaga gatcgtggcc
     1441 aaggaggcac aggtgaaggt ggcagaggtg gagggagagc aggtggacaa taaggccaag
     1501 ctggaggcca ccctgcagga ggaggcagca atccagcagg agcaccggga gaaggagctg
     1561 cagaagagat ctgaggtggc caaggatttt gagcccgaga gggtggtggc agcaccacag
     1621 aggcctggaa ccgagcccca gcctgagatg cctgacacag tgctgcagag cgagaccctg
     1681 aaggatacag ccccagtgct ggagggcctg aaggaggagg agatcaccaa ggaggagatc
     1741 gacatcctgt ctgatgcctg tagcaagctg caggagcaga agaagagcct gacaaaggag
     1801 aaggaggagc tggagctgct gaaggaggac gtgcaggatt actctgagga cctgcaggag
     1861 atcaagaagg agctgagcaa gaccggcgag gagaagtatg tggaggagag caaggcctcc
     1921 aagaggctga caaagcgcgt gcagcagatg atcggccaga tcgacggcct gatcagccag
     1981 ctggagatgg atcagcaggc aggcaagctg gcaccagcaa acggaatgcc aaccggcgag
     2041 aatgtgatct ccgtggccga gctgatcaac gccatgaagc aggtgaagca catccccgag
     2101 tccaagctga catctctggc cgccgccctg gacgagaaca aggatggcaa ggtcaatatc
     2161 gacgatctgg tgaaagtgat cgagctggtg gacaaggagg atgtgcacat cagcacctcc
     2221 caggtggccg agatcgtggc cacactggag aaggaggaga aggtggagga gaaggagaag
     2281 gccaaggaga aggccgagaa ggaggtggcc gaggtgaagt cctgagaccc agctttcttg
     2341 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     2401 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2461 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2521 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2581 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2641 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2701 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2761 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     2821 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     2881 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     2941 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3001 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat
     3061 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3121 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     3181 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3241 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3301 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3361 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3421 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3481 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3541 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3601 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3661 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3721 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3781 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3841 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3901 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3961 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4021 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4081 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4141 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4201 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4261 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4321 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     4381 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc
     4441 gctagccagg aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct
     4501 tagtttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct
     4561 tcacaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca
     4621 aacaacagat aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc
     4681 tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgttgt
     4741 aaaacgacgg ccagtcttaa gctcgggccc
//
