LOCUS       sequence-479345-        2982 bp    DNA     circular SYN 30-JUL-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 2982)
  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 2982)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 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-SLC31A2_STOP
FEATURES             Location/Qualifiers
     source          1..2982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    241..340
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(779..878)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(895..914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(896..914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(919..935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(919..935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1037..1056)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1048..1857
                     /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(1115..1134)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2004..2592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2493..2512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2746..2763
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2922..2949)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
      241 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacacat tgatgagcaa
      301 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccaccatggc catgcacttc
      361 atcttttccg acaccgccgt gctgctgttc gatttttggt ccgtgcactc tccagcagga
      421 atggcactgt ctgtgctggt gctgctgctg ctggccgtgc tgtacgaggg catcaaggtc
      481 ggcaaggcca agctgctgaa ccaggtgctg gtgaatctgc ccaccagcat ctcccagcag
      541 accatcgcag agacagacgg cgatagcgcc ggatctgaca gcttccctgt gggacggaca
      601 caccacagat ggtacctgtg ccactttggc cagtctctga tccacgtgat ccaggtggtc
      661 atcggctact tcatcatgct ggccgtgatg agctataaca cctggatctt tctgggagtg
      721 gtgctgggaa gcgccgtggg atactatctg gcctatcccc tgctgtccac agcctaagac
      781 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
      841 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
      901 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
      961 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1021 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1081 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1141 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1201 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1261 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1321 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1381 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1441 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1501 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1561 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1621 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1681 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     1741 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     1801 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     1861 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     1921 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     1981 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2041 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2101 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2161 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2221 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2281 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2341 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2401 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2461 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2521 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2581 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2641 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2701 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2761 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2821 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2881 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     2941 tggccttctg cttagtttga tgcctggcag tttatggcgg gc
//
