LOCUS       sequence-261922-        5499 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 5499)
  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 5499)
  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_SLC4A9
FEATURES             Location/Qualifiers
     source          1..5499
                     /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(3624..3723)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3740..3759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3741..3759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3764..3780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3764..3780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3882..3901)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3893..4702
                     /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(3960..3979)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4849..5437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5338..5357
                     /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 tgaagctgcc aggacaggag ggattcgagg caagctccgc
      721 cccaaggaac atcccaagcg gagagctgga ctccaatcct gatccaggaa ccggaccatc
      781 ccctgacgga ccttctgata cagagagcaa ggagctgggc gtgcccaagg accctctgct
      841 gtttatccag ctgaacgagc tgctgggatg gccacaggcc ctggagtgga gagagaccgg
      901 atctagctcc gccagcctgc tgctggatat gggcgagatg ccctccatca ccctgtctac
      961 acacctgcat caccggtggg tgctgttcga ggagaagctg gaggtggcag caggcagatg
     1021 gtctgcccca cacgtgccca ccctggccct gccatccctg cagaagctga ggtctctgct
     1081 ggcagaggga ctggtgctgc tggactgccc tgcacagagc ctgctggagc tggtggagca
     1141 ggtgacaagg gtggagtccc tgtctcctga gctgagggga cagctgcagg ccctgctgct
     1201 gcagcgccca cagcactaca atcagaccac aggaaccagg ccatgctggg gaagcacaca
     1261 ccccagaaag gcatccgata acgaggaggc accactgagg gagcagtgtc agaatcctct
     1321 gagacagaag ctgccccctg gagcagaggc aggaaccgtg ctggcaggag agctgggctt
     1381 tctggcacag cctctgggag ccttcgtgag gctgaggaac ccagtggtgc tgggatccct
     1441 gacagaggtg agcctgccct cccggttctt ttgcctgctg ctgggccctt gtatgctggg
     1501 caagggctat cacgagatgg gaagagcagc agccgtgctg ctgtctgacc cacagttcca
     1561 gtggtctgtg cggagagcca gcaatctgca cgacctgctg gccgccctgg atgcctttct
     1621 ggaggaggtg accgtgctgc caccaggccg gtgggatccc acagccagaa tccctccacc
     1681 caagtgtctg ccttctcagc acaagaggct gccaagccag cagagggaga tcagaggacc
     1741 tgcagtgcca aggctgacca gcgccgagga caggcacaga cacggaccac acgcacactc
     1801 cccagagctg cagaggacag gccgcctgtt cggaggactg atccaggatg tgaggcgcaa
     1861 ggtgccctgg tacccttccg actttctgga tgccctgcac ctgcagtgct tctctgccgt
     1921 gctgtacatc tatctggcca ccgtgacaaa cgccatcacc tttggcggac tgctgggcga
     1981 cgcaacagat ggagcccagg gcgtgctgga gtcctttctg ggaaccgcag tggcaggagc
     2041 agcattctgt ctgatggccg gccagccact gaccatcctg tctagcacag gccccgtgct
     2101 ggtgttcgag aggctgctgt ttagcttctc ccgcgactac agcctggatt atctgccctt
     2161 taggctgtgg gtgggcatct gggtggccac cttctgcctg gtgctggtgg caacagaggc
     2221 aagcgtgctg gtgaggtact tcacccgctt cacagaggag ggcttttgtg ccctgatctc
     2281 cctgatcttc atctacgacg ccgtgggcaa gatgctgaat ctgacccaca catatcctat
     2341 ccagaagcca ggctcctctg cctacggatg cctgtgccag tatccaggac caggaggaaa
     2401 cgagagccag tggatcagga cccgccccaa ggacagagac gatatcgtga gcatggatct
     2461 gggcctgatc aatgcatccc tgctgcctcc accagagtgc accaggcagg gaggacaccc
     2521 aaggggacca ggctgtcaca cagtgcctga tatcgccttc ttttccctgc tgctgttcct
     2581 gacctctttc tttttcgcca tggccctgaa gtgcgtgaag acatcccggt ttttcccatc
     2641 tgtggtgaga aagggcctga gcgactttag ctccgtgctg gcaatcctgc tgggatgtgg
     2701 actggatgcc tttctgggcc tggccacccc taagctgatg gtgccaaggg agttcaagcc
     2761 tacactgcca ggaaggggat ggctggtgtc tccctttggc gccaaccctt ggtggtggag
     2821 cgtggcagcc gccctgcccg ccctgctgct gtccatcctg atcttcatgg accagcagat
     2881 caccgccgtg atcctgaatc ggatggagta cagactgcag aagggcgccg gctttcacct
     2941 ggatctgttc tgcgtggccg tgctgatgct gctgaccagc gccctgggac tgccttggta
     3001 tgtgagcgcc acagtgatct ccctggccca catggactct ctgcggagag agagcagggc
     3061 atgtgcacca ggagagaggc ctaacttcct gggaatccgg gagcagagac tgaccggcct
     3121 ggtggtgttt atcctgacag gcgcctccat ctttctggcc cctgtgctga agttcatccc
     3181 tatgccagtg ctgtacggca tctttctgta tatgggcgtg gccgccctgt ctagcatcca
     3241 gttcaccaat agggtgaagc tgctgctgat gccagccaag caccagccag acctgctgct
     3301 gctgaggcac gtgccactga ccagagtgca cctgttcaca gcaatccagc tggcatgcct
     3361 gggactgctg tggatcatca agagcacacc tgccgccatc atcttcccac tgatgctgct
     3421 gggactggtg ggcgtgagga aggccctgga gcgcgtgttt tccccacagg agctgctgtg
     3481 gctggatgag ctgatgcccg aggaggagcg gagcatccca gagaagggac tggagcctga
     3541 gcactctttt agcggctccg actctgagga ttctgagctg atgtaccagc ccaaggcccc
     3601 tgagatcaac atcagcgtga atgacccagc tttcttgtac aaagttggca ttataagaaa
     3661 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     3721 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3781 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3841 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3901 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3961 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     4021 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     4081 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     4141 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     4201 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     4261 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     4321 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     4381 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     4441 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     4501 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4561 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4621 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     4681 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     4741 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     4801 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4981 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     5401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     5461 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
