LOCUS       sequence-458142-        4329 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 4329)
  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 4329)
  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-SLC13A2_STOP
FEATURES             Location/Qualifiers
     source          1..4329
                     /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"
     protein_bind    complement(2454..2553)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2570..2589)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2571..2589)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2594..2610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2594..2610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2712..2731)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2723..3532
                     /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(2790..2809)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3679..4267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4168..4187
                     /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 accatggcaa cctgctggca ggccctgtgg gcataccggt cttatctgat
      721 cgtgttcttt gtgcctatcc tgctgctgcc actgcccatc ctggtgccaa gcaaggaggc
      781 ctactgcgcc tatgccatca tcctgatggc cctgttctgg tgtacagagg ccctgccact
      841 ggcagtgacc gccctgtttc ctctgatcct gttcccaatg atgggaatcg tggacgcatc
      901 cgaggtggca gtggagtacc tgaaggattc taacctgctg ttctttggag gactgctggt
      961 ggcaatcgca gtggagcact ggaatctgca caagcggatc gccctgagag tgctgctgat
     1021 cgtgggcgtg cggcctgcac cactgatcct gggctttatg ctggtgaccg ccttcctgag
     1081 catgtggatc tccaacaccg ccacatccgc catgatggtg ccaattgctc acgctgttct
     1141 ggatcagctg cacagctccc aggcctctag caatgtggag gagggctcta acaatcccac
     1201 cttcgagctg caggagccca gccctcagaa ggaggtgaca aagctggata acggacaggc
     1261 cctgcctgtg accagcgcct cctctgaggg aagagcacac ctgtcccaga agcacctgca
     1321 cctgacacag tgcatgtccc tgtgcgtgtg ctacagcgcc tccatcggag gaatcgccac
     1381 cctgacagga accgcaccaa acctggtgct gcagggacag atcaattctc tgttccccca
     1441 gaacggcaat gtggtgaact ttgcctcttg gttcagcttt gccttcccta ccatggtcat
     1501 cctgctgctg ctggcctggc tgtggctgca gatcctgttc ctgggcttta acttcaggaa
     1561 gaattttggc atcggcgaga agatgcagga gcagcagcag gccgcctatt gcgtgatcca
     1621 gacagagcac cgcctgctgg gcccaatgac cttcgccgag aaggccatct ccatcctgtt
     1681 cgtgatcctg gtgctgctgt ggtttacaag ggagcccggc ttctttctgg gctggggcaa
     1741 cctggccttt cctaatgcca agggcgagtc catggtgtct gacggcacag tggccatctt
     1801 tatcggcatc atcatgttca tcatcccaag caagtttccc ggcctgaccc aggaccccga
     1861 gaatcctggc aagctgaagg cccctctggg cctgctggat tggaagacag tgaaccagaa
     1921 gatgccatgg aatatcgtgc tgctgctggg aggaggatac gcactggcca agggaagcga
     1981 gcggagcgga ctgtctgagt ggctgggcaa taagctgacc ccactgcagt ccgtgccagc
     2041 accagcaatc gcaatcatcc tgtctctgct ggtggccaca ttcaccgagt gcacaagcaa
     2101 cgtggccacc acaaccatct ttctgccaat cctggccagc atggcacagg caatctgcct
     2161 gcacccactg tatgtgatgc tgccttgtac actggccacc tccctggcct tcatgctgcc
     2221 agtggcaacc ccacctaacg ccatcgtgtt ttctttcggc gacctgaagg tgctggatat
     2281 ggcccgggcc ggctttctgc tgaacatcat cggcgtgctg atcatcgccc tggccatcaa
     2341 tagctggggc atcccactgt ttagcctgca ctccttcccc tcttgggccc agtccaacac
     2401 aaccgcccag tgtctgccct ctctggccaa tacaaccaca cctagcccat aagacccagc
     2461 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     2521 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     2581 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2641 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2701 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2761 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2821 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2881 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     2941 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3001 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3061 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3121 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3181 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3241 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3301 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3361 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3421 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3481 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3541 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3601 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3661 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3721 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3781 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3841 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3901 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3961 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4021 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4081 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4141 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4201 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4261 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4321 tcacatgtt
//
