LOCUS       sequence-263896-        4140 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 4140)
  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 4140)
  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_SLC45A2
FEATURES             Location/Qualifiers
     source          1..4140
                     /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(2265..2364)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2381..2400)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2382..2400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2405..2421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2405..2421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2523..2542)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2534..3343
                     /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(2601..2620)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3490..4078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3979..3998
                     /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 accatgggct ctaacagcgg ccaggccggc agacacatct acaagagcct
      721 ggccgacgat ggcccattcg actccgtgga gccccctaag agacccacct ccaggctgat
      781 catgcactct atggccatgt ttggcaggga gttctgctac gccgtggagg ccgcctatgt
      841 gacacctgtg ctgctgagcg tgggcctgcc aagctccctg tatagcatcg tgtggtttct
      901 gtcccccatc ctgggcttcc tgctgcagcc tgtggtggga tccgcctctg atcactgtcg
      961 ctccagatgg ggccggagaa ggccctacat cctgaccctg ggcgtgatga tgctggtggg
     1021 catggccctg tatctgaacg gagcaacagt ggtggccgcc ctgatcgcaa atccaaggcg
     1081 gaagctggtg tgggccatct ccgtgaccat gatcggcgtg gtgctgtttg acttcgccgc
     1141 cgactttatc gatggcccca tcaaggccta cctgttcgac gtgtgctctc accaggataa
     1201 ggagaagggc ctgcactacc acgcactgtt taccggattc ggaggcgccc tgggatatct
     1261 gctgggagcc atcgattggg cacacctgga gctgggccgg ctgctgggca cagagtttca
     1321 ggtcatgttc tttttcagcg ccctggtgct gaccctgtgc ttcacagtgc acctgtgcag
     1381 catctccgag gcacctctga ccgaggtggc aaagggaatc ccaccacagc agacaccaca
     1441 ggaccctcca ctgtctagcg atggcatgta cgagtatggc tctatcgaga aggtgaagaa
     1501 cggctacgtg aatcctgagc tggccatgca gggcgccaag aacaagaatc acgccgagca
     1561 gaccagaagg gccatgacac tgaagtccct gctgagagcc ctggtgaaca tgccccctca
     1621 ctacaggtat ctgtgcatct ctcacctgat cggctggacc gcctttctga gcaatatgct
     1681 gtttttcaca gacttcatgg gccagatcgt gtaccgcggc gatccatatt ctgcccacaa
     1741 cagcaccgag tttctgatct atgagagggg agtggaagtg ggatgctggg gactgtgcat
     1801 caattccgtg ttttcctctc tgtactctta tttccagaag gtgctggtga gctacatcgg
     1861 cctgaagggc ctgtacttta ccggctatct gctgttcggc ctgggcacag gctttatcgg
     1921 cctgttcccc aacgtgtata gcaccctggt gctgtgcagc ctgtttggcg tgatgagctc
     1981 caccctgtac acagtgcctt tcaacctgat cacagagtat cacagggagg aggagaagga
     2041 gaggcagcag gcaccaggag gcgacccaga taatagcgtg cgcggcaagg gaatggactg
     2101 cgccaccctg acatgtatgg tgcagctggc acagatcctg gtgggaggag gactgggctt
     2161 cctggtgaat accgccggca cagtggtggt ggtggtcatc accgcatccg ccgtggccct
     2221 gatcggatgc tgttttgtgg ccctgttcgt gcggtacgtg gatgacccag ctttcttgta
     2281 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     2341 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     2401 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     2461 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     2521 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa
     2581 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     2641 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     2701 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt
     2761 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     2821 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     2881 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     2941 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     3001 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     3061 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga
     3121 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     3181 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     3241 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     3301 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg
     3361 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa
     3421 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3481 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3541 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3601 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3661 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3721 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3781 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3841 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3901 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3961 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4021 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4081 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
