LOCUS       sequence-355839-        4200 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 4200)
  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 4200)
  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-SPNS2_STOP
FEATURES             Location/Qualifiers
     source          1..4200
                     /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(1757..1856)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1873..1892)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1874..1892)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1897..1913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1897..1913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2015..2034)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2026..2835
                     /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(2093..2112)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2982..3570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3471..3490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3724..3741
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3900..3927)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4019..4105)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4154..4176
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4168..4185
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4169..4185
                     /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 ccaccatgat gtgcctggag
      121 tgtgcatccg ccgcagcagg aggagcagag gaggaggagg ccgacgccga gcggagaagg
      181 cgccggagag gcgcccagag gggcgccggc ggcagcggct gctgtggagc aaggggagca
      241 ggaggagcag gcgtgtccgc cgccggcgat gaggtgcaga ccctgagcgg ctccgtgagg
      301 cgcgccccaa caggcccccc tggcacccca ggcacacccg gctgtgcagc aaccgcaaag
      361 ggaccaggag cacagcagcc caagcctgcc tctctgggaa ggggaagggg agcagcagca
      421 gcaatcctga gcctgggcaa cgtgctgaat tacctggacc ggtatacagt ggccggcgtg
      481 ctgctggaca tccagcagca cttcggcgtg aaggatagag gcgccggcct gctgcagtcc
      541 gtgttcatct gttcttttat ggtggccgcc ccaatcttcg gctacctggg cgatcggttt
      601 aacagaaaag tgatcctgag ctgcggcatc ttcttttggt ccgccgtgac cttcagctcc
      661 tcttttatcc cccagcagta cttctggctg ctggtgctgt ccaggggact ggtgggaatc
      721 ggagaggcct cttatagcac catcgccccc acaatcatcg gcgacctgtt taccaagaat
      781 accagaacac tgatgctgag cgtgttctac tttgccatcc ctctgggctc cggcctgggc
      841 tatatcacag gaagctccgt gaagcaggca gcaggcgatt ggcactgggc actgagggtg
      901 agccctgtgc tgggcatgat caccggcaca ctgatcctga tcctggtgcc agccaccaag
      961 aggggacacg cagaccagct gggcgatcag ctgaaggccc ggacatcctg gctgagagac
     1021 atgaaggccc tgatccggaa cagatcttac gtgttctcta gcctggccac ctccgccgtg
     1081 tcttttgcca caggcgccct gggcatgtgg atccctctgt atctgcacag ggcccaggtg
     1141 gtgcagaaga ccgccgagac atgcaattct ccaccctgtg gcgccaagga cagcctgatc
     1201 ttcggcgcca tcacctgttt cacaggcttt ctgggcgtgg tgaccggagc aggagcaaca
     1261 aggtggtgcc gcctgaagac ccagagggca gatccactgg tgtgcgcagt gggaatgctg
     1321 ggatccgcca tcttcatctg cctgatcttt gtggccgcca agtcctctat cgtgggcgcc
     1381 tacatctgca tcttcgtggg cgagaccctg ctgttttcta actgggccat cacagccgac
     1441 atcctgatgt atgtggtcat ccctaccagg agagcaacag cagtggccct gcagagcttc
     1501 acctcccacc tgctgggcga tgcaggatcc ccatacctga tcggctttat ctctgacctg
     1561 atcaggcagt ctacaaagga tagccccctg tgggagttcc tgagcctggg ctatgccctg
     1621 atgctgtgcc cttttgtggt ggtgctgggc ggcatgttct ttctggccac cgccctgttc
     1681 ttcgtgagcg atagggcaag ggcagagcag caagtgaatc agctggccat gcctccagcc
     1741 agcgtgaagg tgtaagaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1801 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1861 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1921 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1981 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     2041 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     2101 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     2161 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     2221 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     2281 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     2341 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2401 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2461 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2521 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2581 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2641 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2701 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2761 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2821 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2881 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2941 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3001 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3061 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3121 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3181 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3241 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3301 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3361 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3421 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3481 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3541 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3601 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3661 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     3721 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     3781 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3841 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     3901 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     3961 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     4021 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     4081 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     4141 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
//
