LOCUS       sequence-262936-        4524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4524)
  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 4524)
  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_SLC28A2
FEATURES             Location/Qualifiers
     source          1..4524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2106..2115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2649..2748)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2765..2784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2766..2784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2789..2805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2789..2805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2907..2926)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2918..3727
                     /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(2985..3004)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3874..4462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4363..4382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc accatggaga aggcctctgg caggcagagc atcgccctgt ccaccgtgga
      721 gaccggcaca gtgaacccag gcctggagct gatggagaag gaggtggagc ccgagggcag
      781 caagaggacc gacgcacagg gacactccct gggcgatgga ctgggaccat ctacatacca
      841 gcggagaagc aggtggcctt tctctaaggc ccgcagcttt tgcaagaccc acgcctccct
      901 gttcaagaag atcctgctgg gactgctgtg cctggcatac gcagcatatc tgctggcagc
      961 atgtatcctg aacttccagc gggccctggc cctgtttgtg atcacctgcc tggtcatctt
     1021 cgtgctggtg cacagctttc tgaagaagct gctgggcaag aagctgacca gatgtctgaa
     1081 gcccttcgag aatagcaggc tgcgcctgtg gacaaagtgg gtgtttgcag gcgtgtccct
     1141 ggtgggactg atcctgtggc tggccctgga caccgcacag aggccagagc agctgatccc
     1201 cttcgccggc atctgcatgt tcatcctgat cctgtttgcc tgttccaagc accactctgc
     1261 cgtgagctgg cggacagtgt tttctggcct gggcctgcag ttcgtgtttg gcatcctggt
     1321 catcagaacc gatctgggct acacagtgtt ccagtggctg ggcgagcagg tgcagatctt
     1381 tctgaactat accgtggccg gcagctcctt cgtgtttggc gacacactgg tgaaggacgt
     1441 gttcgccttt caggccctgc ctatcatcat cttctttggc tgcgtggtgt ccatcctgta
     1501 ctatctgggc ctggtgcagt gggtggtgca gaaggtggcc tggttcctgc agatcaccat
     1561 gggcaccaca gcaaccgaga cactggcagt ggcaggcaat atctttgtgg gcatgacaga
     1621 ggcccctctg ctgatccggc catacctggg cgacatgacc ctgtctgaga tccacgccgt
     1681 gatgacaggc ggcttcgcaa ccatcagcgg aacagtgctg ggagccttca tcgcctttgg
     1741 cgtggatgcc tctagcctga tctctgccag cgtgatggca gcaccttgtg cactggcatc
     1801 ctctaagctg gcctatccag aggtggagga gtccaagttt aagtctgagg agggagtgaa
     1861 gctgccaagg ggcaaggaga gaaacgtgct ggaggcagca agcaatggag cagtggacgc
     1921 aatcggactg gcaaccaacg tggcagccaa tctgatcgcc ttcctggccg tgctggcctt
     1981 tatcaacgcc gccctgtcct ggctgggaga gctggtggac atccagggac tgaccttcca
     2041 ggtcatctgc tcttacctgc tgcggccaat ggtgtttatg atgggcgtgg agtggacaga
     2101 ttgtcccatg gtggccgaga tggtgggcat caagttcttt atcaacgagt tcgtggccta
     2161 ccagcagctg tcccagtata agaataagag gctgtctggc atggaggagt ggatcgaggg
     2221 cgagaagcag tggatctctg tgcgcgccga gatcatcacc acattcagcc tgtgcggctt
     2281 tgccaatctg agctccatcg gcatcaccct gggcggcctg acatccatcg tgcctcacag
     2341 gaagtccgac ctgtctaagg tggtggtgcg cgccctgttc accggagcat gcgtgagcct
     2401 gatctccgcc tgtatggcag gaatcctgta cgtgccaaga ggagcagagg cagattgcgt
     2461 gagcttccct aacacctcct ttacaaatcg gacctacgag acatatatgt gctgtagagg
     2521 cctgttccag tccacctctc tgaacggcac aaatccccct agcttctccg gcccatggga
     2581 ggataaggag tttagcgcca tggccctgac caattgctgt ggcttttata acaatacagt
     2641 gtgcgccgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2701 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2761 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2821 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2881 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2941 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3001 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3061 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3121 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3181 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3241 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3301 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3361 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3421 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3481 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3541 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3601 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3661 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3721 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3781 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4021 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4381 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4441 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4501 ttgctggcct tttgctcaca tgtt
//
