LOCUS       sequence-261076-        4500 bp    DNA     circular SYN 14-APR-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 4500)
  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 4500)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SLC23A2
FEATURES             Location/Qualifiers
     source          1..4500
                     /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(2625..2724)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2741..2760)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2742..2760)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2765..2781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2765..2781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2883..2902)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2894..3703
                     /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(2961..2980)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3850..4438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4339..4358
                     /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 accatgatgg gcatcggcaa gaacaccaca agcaagtcca tggaggcagg
      721 aagctccacc gagggcaagt acgaggacga ggccaagcac ccagccttct ttacactgcc
      781 agtggtcatc aacggaggag ccacctctag cggagagcag gacaatgagg atacagagct
      841 gatggccatc tataccacag agaatggcat cgccgagaag tcctctctgg ccgagacact
      901 ggactctacc ggaagcctgg acccacagag gtccgatatg atctacacca tcgaggatgt
      961 gcccccttgg tatctgtgca tcttcctggg cctgcagcac tacctgacat gtttctctgg
     1021 caccatcgcc gtgccttttc tgctggccga cgccatgtgc gtgggctatg atcagtgggc
     1081 cacaagccag ctgatcggca ccatcttctt ttgcgtgggc atcaccacac tgctgcagac
     1141 cacattcggc tgtcggctgc ccctgtttca ggcaagcgcc ttcgcctttc tggcacctgc
     1201 cagagccatc ctgagcctgg acaagtggaa gtgtaacacc acagacgtga gcgtggccaa
     1261 tggcacagcc gagctgctgc acaccgagca catctggtac cctaggatcc gcgagatcca
     1321 gggcgccatc atcatgagct ccctgatcga ggtggtcatc ggactgctgg gactgccagg
     1381 cgccctgctg aagtatatcg gccccctgac catcacacct accgtggccc tgatcggact
     1441 gtccggattc caggcagcag gagagagggc aggcaagcac tggggcatcg ccatgctgac
     1501 catcttcctg gtgctgctgt tttcccagta cgccagaaac gtgaagtttc cactgcccat
     1561 ctacaagtct aagaagggct ggacagccta taagctgcag ctgttcaaga tgtttccaat
     1621 catcctggcc atcctggtga gctggctgct gtgcttcatc tttacagtga ccgacgtgtt
     1681 cccacccgat tccacaaagt acggctttta tgccaggacc gatgcaaggc agggcgtgct
     1741 gctggtggca ccatggttca aggtgcctta tccatttcag tggggactgc caaccgtgtc
     1801 tgccgcaggc gtgatcggaa tgctgagcgc cgtggtggcc agcatcatcg agtccatcgg
     1861 cgactactat gcctgcgcaa ggctgagctg tgcacctcca ccacctatcc acgccatcaa
     1921 ccgcggcatc ttcgtggagg gcctgtcttg cgtgctggat ggcatctttg gcacaggcaa
     1981 cggcagcacc tctagctccc ctaatatcgg cgtgctgggc atcaccaaag tgggctccag
     2041 gagagtgatc cagtgcggag ccgccctgat gctggccctg ggcatgatcg gcaagttctc
     2101 tgccctgttt gcaagcctgc cagaccctgt gctgggcgcc ctgttctgta cactgtttgg
     2161 catgatcacc gccgtgggcc tgtctaatct gcagttcatc gatctgaact ctagcaggaa
     2221 tctgttcgtg ctgggctttt ccatcttctt tggcctggtg ctgccttctt acctgcgcca
     2281 gaacccactg gtgacaggca tcaccggcat cgaccaggtg ctgaatgtgc tgctgaccac
     2341 agccatgttc gtgggcggct gcgtggcctt tatcctggat aacacaatcc caggaacccc
     2401 agaggagagg ggaatcagaa agtggaagaa gggcgtgggc aagggcaata agagcctgga
     2461 cggcatggag tcctacaacc tgcccttcgg catgaatatc atcaagaagt acagatgttt
     2521 ctcctatctg cctatctctc caacatttgt gggctatacc tggaagggcc tgcggaagtc
     2581 cgataattct agatcctctg acgaggatag ccaggccacc ggcgacccag ctttcttgta
     2641 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2701 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2761 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2821 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2881 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2941 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     3001 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     3061 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     3121 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     3181 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     3241 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     3301 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3361 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3421 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3481 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3541 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3601 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3661 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3721 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3781 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3841 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3901 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3961 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4021 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4081 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4141 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4201 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4261 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4321 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4381 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4441 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
