LOCUS       sequence-259549-        4584 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 4584)
  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 4584)
  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_SLC25A12
FEATURES             Location/Qualifiers
     source          1..4584
                     /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"
     CDS             1545..1559
                     /codon_start=1
                     /product="tag for protein cleavage using Ni2+ ion (Dang et 
                     al., 2019)"
                     /label=SNAC tag
                     /translation="GSHHW"
     protein_bind    complement(2709..2808)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2825..2844)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2826..2844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2849..2865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2849..2865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2967..2986)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2978..3787
                     /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(3045..3064)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3934..4522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4423..4442
                     /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 accatggccg tgaaggtgca gaccacaaag aggggcgacc cacacgagct
      721 gcgcaacatc ttcctgcagt acgccagcac cgaggtggac ggcgagaggt atatgacacc
      781 cgaggatttt gtgcagcggt acctgggcct gtataatgat cccaactcca atcctaagat
      841 cgtgcagctg ctggcaggag tggcagacca gaccaaggat ggcctgatct cttaccagga
      901 gttcctggcc tttgagtccg tgctgtgcgc ccctgactct atgttcatcg tggccttcca
      961 gctgtttgat aagtccggca acggcgaggt gacctttgag aatgtgaagg agatcttcgg
     1021 ccagacaatc atccaccacc acatcccctt caactgggac tgtgagttta tccggctgca
     1081 cttcggccac aacagaaaga agcacctgaa ttataccgag ttcacacagt ttctgcagga
     1141 gctgcagctg gagcacgcaa ggcaggcctt cgccctgaag gataagtcta agagcggcat
     1201 gatcagcggc ctggacttct ccgatatcat ggtgaccatc agatctcaca tgctgacacc
     1261 ctttgtggag gagaatctgg tgagcgccgc aggaggatcc atctctcacc aggtgagctt
     1321 ctcctacttt aacgccttca attccctgct gaacaatatg gagctggtgc ggaagatcta
     1381 ttctaccctg gccggcacaa gaaaggacgt ggaggtgacc aaggaggagt tcgcacagag
     1441 cgccatcaga tacggacagg tgacacctct ggagatcgac atcctgtacc agctggccga
     1501 tctgtataac gcctccggca ggctgaccct ggcagatatc gagaggatcg caccactggc
     1561 agagggcgcc ctgccttata atctggcaga gctgcagagg cagcagtccc caggactggg
     1621 cagacccatc tggctgcaga tcgcagagag cgcctaccgg ttcaccctgg gctccgtggc
     1681 aggagcagtg ggagcaaccg ccgtgtatcc catcgacctg gtgaagacaa ggatgcagaa
     1741 ccagagggga tctggaagcg tggtgggaga gctgatgtac aagaattctt ttgattgctt
     1801 caagaaggtg ctgcggtacg agggcttctt tggcctgtat agaggcctga tcccacagct
     1861 gatcggagtg gcaccagaga aggccatcaa gctgaccgtg aacgactttg tgagggataa
     1921 gttcacacgg agagacggca gcgtgccact gccagcagag gtgctggcag gaggctgtgc
     1981 cggcggctcc caggtcatct tcaccaaccc tctggagatc gtgaagatca ggctgcaggt
     2041 ggcaggagag atcaccacag gaccaagggt gtctgccctg aatgtgctgc gcgacctggg
     2101 catcttcggc ctgtacaagg gcgccaaggc ctgctttctg cgcgatatcc cttttagcgc
     2161 catctacttc ccagtgtatg cccactgtaa gctgctgctg gcagatgaga acggacacgt
     2221 gggaggcctg aatctgctgg cagcaggagc aatggcagga gtgccagccg ccagcctggt
     2281 gacccctgcc gacgtgatca agacaaggct gcaggtggca gcaagagcag gacagaccac
     2341 atactctggc gtgatcgatt gcttccggaa gatcctgaga gaggagggcc caagcgcctt
     2401 ttggaaggga accgcagcaa gggtgtttcg cagctcccca cagttcggag tgaccctggt
     2461 gacatacgag ctgctgcagc ggtggtttta tatcgacttc ggaggcctga agccagcagg
     2521 atctgagcct acaccaaaga gcaggatcgc agacctgcca cctgcaaacc ctgatcacat
     2581 cggaggatac aggctggcaa ccgccacatt tgccggcatc gagaataagt tcggcctgta
     2641 tctgcctaag tttaagtccc catctgtggc cgtggtgcag cccaaggccg ccgtggccgc
     2701 cacccaggac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2761 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2821 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2881 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2941 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     3001 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     3061 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     3121 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     3181 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     3241 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     3301 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3361 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3421 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3481 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3541 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3601 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3661 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3721 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3781 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3841 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4561 ttgctggcct tttgctcaca tgtt
//
