LOCUS       sequence-351782-        3522 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 3522)
  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 3522)
  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-SLC25A27_STOP
FEATURES             Location/Qualifiers
     source          1..3522
                     /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(1079..1178)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1195..1214)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1196..1214)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1219..1235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1219..1235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1337..1356)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1348..2157
                     /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(1415..1434)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2304..2892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2793..2812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3046..3063
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3222..3249)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3341..3427)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3476..3498
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3490..3507
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3491..3507
                     /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 ccaccatgtc tgtgcccgag
      121 gaggaggaga gactgctgcc actgacccag aggtggccta gggccagcaa gttcctgctg
      181 tccggatgcg cagcaaccgt ggcagagctg gccacatttc ctctggacct gaccaagaca
      241 aggctgcaga tgcagggaga ggccgccctg gcaaggctgg gcgatggagc aagagagagc
      301 gccccataca ggggaatggt gagaacagcc ctgggcatca tcgaggagga gggcttcctg
      361 aagctgtggc agggagtgac cccagcaatc tacaggcacg tggtgtattc cggcggccgc
      421 atggtgacat acgagcacct gcgggaggtg gtgttcggca agtccgagga cgagcactat
      481 ccactgtgga agtctgtgat cggaggaatg atggcaggcg tgatcggaca gtttctggcc
      541 aaccccaccg atctggtgaa ggtgcagatg cagatggagg gcaagcgcaa gctggagggc
      601 aagcctctgc ggttcagagg cgtgcaccac gcctttgcca agatcctggc agagggagga
      661 atcagaggac tgtgggccgg ctgggtgcct aacatccaga gggccgccct ggtgaatatg
      721 ggcgacctga ccacatacga taccgtgaag cactatctgg tgctgaacac cccactggag
      781 gacaatatca tgacacacgg cctgagctcc ctgtgctctg gactggtggc aagcatcctg
      841 ggaacccctg ccgacgtgat caagagccgg atcatgaatc agccaaggga taagcagggc
      901 cgcggcctgc tgtacaagtc tagcacagat tgtctgatcc aggccgtgca gggcgagggc
      961 ttcatgtccc tgtataaggg ctttctgcca tcttggctgc gcatgacccc ctggagcatg
     1021 gtgttctggc tgacatatga gaagatccgg gagatgtccg gcgtgtctcc tttttaagac
     1081 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     1141 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     1201 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     1261 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     1321 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     1381 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     1441 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     1501 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     1561 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     1621 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     1681 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     1741 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     1801 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     1861 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     1921 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     1981 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2041 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2101 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2161 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     2221 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2281 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2341 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2401 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2461 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2521 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2581 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2641 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2701 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2761 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2821 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2881 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2941 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3001 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3061 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3121 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3181 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3241 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3301 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3361 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3421 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     3481 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
