LOCUS       sequence-263914-        6081 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 6081)
  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 6081)
  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_SLC9C1
FEATURES             Location/Qualifiers
     source          1..6081
                     /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(4206..4305)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4322..4341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4323..4341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4346..4362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4346..4362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4464..4483)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4475..5284
                     /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(4542..4561)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /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 gcatcttcaa ggagttcttt ttctctaccg aggacctgcc
      721 tgaagtgatc ctgaccctga gcctgatcag ctccatcggc gccttcctga atcggcacct
      781 ggaggatttt cccatccctg tgccagtgat cctgtttctg ctgggctgta gcttcgaggt
      841 gctgtccttt acctctagcc aggtgcagag gtacgccaac gccatccagt ggatgtcccc
      901 cgacctgttt ttccgcatct tcacccctgt ggtgtttttc accacagcct ttgacatgga
      961 tacatacatg ctgcagaagc tgttctggca gatcctgctg atcagcatcc caggctttct
     1021 ggtgaattat atcctggtgc tgtggcacct ggcctccgtg aaccagctgc tgctgaagcc
     1081 cacccagtgg ctgctgttct ctgccatcct ggtgtcctct gaccctatgc tgacagcagc
     1141 agcaatcagg gatctgggac tgtccagatc tctgatcagc ctgatcaatg gcgagtccct
     1201 gatgacctct gtgatcagcc tgatcacctt cacaagcatc atggactttg atcagaggct
     1261 gcagtccaag cgcaaccaca cactggccga ggagatcgtg ggcggcatct gctcttacat
     1321 catcgccagc ttcctgtttg gcatcctgag ctccaagctg atccagttct ggatgtccac
     1381 cgtgtttggc gacgatgtga atcacatctc tctgatcttc agcatcctgt acctgatctt
     1441 ttatatctgt gagctggtgg gcatgtctgg catcttcacc ctggccatcg tgggcctgct
     1501 gctgaacagc acatccttta aggccgccat cgaggagacc ctgctgctgg agttctggac
     1561 atttctgtcc cggatcgcct tcctgatggt gtttaccttt ttcggcctgc tgatccccgc
     1621 ccacacatac ctgtatatcg agttcgtgga tatctactat agcctgaata tctatctgac
     1681 cctgatcgtg ctgagatttc tgacactgct gctgatctcc cccgtgctgt ctagggtggg
     1741 ccacgagttc tcttggcgct ggatctttat catggtgtgc agcgagatga agggcatgcc
     1801 taacatcaat atggccctgc tgctggccta ctccgacctg tatttcggct ccgataagga
     1861 gaagtctcag atcctgtttc acggcgtgct ggtgtgcctg atcaccctgg tggtgaacag
     1921 gttcatcctg cctgtggcag tgacaatcct gggactgagg gacgccacca gcacaaagta
     1981 caagtccgtg tgctgtacct tccagcactt tcaggagctg acaaagtccg ccgcctctgc
     2041 cctgaagttt gacaaggatc tggccaacgc cgattggaat atgatcgaga aggccatcac
     2101 cctggagaac ccttatatgc tgaatgagga ggagaccaca gagcaccaga aggtgaagtg
     2161 cccacactgt aacaaggaga tcgacgagat cttcaatacc gaggccatgg agctggccaa
     2221 ccggagactg ctgtctgccc agatcgccag ctaccagcgg cagtatagaa atgagatcct
     2281 gtctcagagc gccgtgcagg tgctggtggg agcagcagag tccttcggcg agaagaaggg
     2341 caagtgcatg tctctggaca ccatcaagaa ctactccgag tctcagaaga ccgtgacatt
     2401 tgcccggaag ctgctgctga actgggtgta caatacaaga aaggagaagg agggcccaag
     2461 caagtacttc ttcttcagga tctgtcacac catcgtgttc acagaggagt ttgagcacgt
     2521 gggctacctg gtcatcctga tgaatatctt cccctttatc atcagctgga tctcccagct
     2581 gaatgtgatc tatcactccg agctgaagca caccaactac tgcttcctga cactgtatat
     2641 cctggaggcc ctgctgaaga tcgccgccat gcgcaaggac ttctttagcc acgcctggaa
     2701 tatctttgag ctggccatca ccctgatcgg catcctgcac gtgatcctga tcgagatcga
     2761 taccatcaag tacatcttca acgagacaga agtgatcgtg ttcatcaagg tggtgcagtt
     2821 ctttaggatc ctgcgcatct tcaagctgat cgccccaaag ctgctgcaga tcatcgataa
     2881 gaggatgtcc caccagaaga ccttttggta cggcatcctg aagggctatg tgcagggcga
     2941 ggccgacatc atgaccatca tcgatcagat cacatctagc aagcagatca agcagatgct
     3001 gctgaagcag gtcatccgca acatggagca cgccatcaag gagctgggct acctggagta
     3061 tgaccacccc gagatcgccg tgaccgtgaa gacaaaggag gagatcaacg tgatgctgaa
     3121 tatggccacc gagatcctga aggccttcgg cctgaagggc atcatcagca agacagaggg
     3181 cgccggcatc aataagctga tcatggccaa gaagaaggag gtgctggata gccagtccat
     3241 catcagacca ctgaccgtgg aggaggtgct gtaccacatc ccctggctgg acaagaacaa
     3301 ggattatatc aatttcatcc aggagaaggc caaggtggtg acattcgact gcggcaacga
     3361 tatctttgag gagggcgacg agcctaaggg catctacatc atcatctctg gcatggtgaa
     3421 gctggagaag agcaagccag gcctgggcat cgaccagatg gtggagagca aggagaagga
     3481 tttccccatc atcgacaccg attatatgct gtccggcgag atcatcggcg agatcaactg
     3541 cctgacaaat gagccaatga agtactccgc cacctgcaag acagtggtgg agacctgttt
     3601 catccctaag acacacctgt atgatgcctt tgagcagtgt tctccactga tcaagcagaa
     3661 gatgtggctg aagctgggac tggcaatcac cgcacggaag atcagagagc acctgagcta
     3721 cgaggactgg aactataata tgcagctgaa gctgagcaat atctacgtgg tggatatccc
     3781 catgtccacc aagacagaca tctacgatga gaacctgatc tatgtgatcc tgatccacgg
     3841 agcagtggag gactgcctgc tgcggaagac ctatagagcc ccattcctga tccccatcac
     3901 atgtcaccag atccagagca tcgaggactt caccaaggtg gtcatcatcc agacccctat
     3961 caacatgaag acattcaggc gcaatatccg gaagtttgtg cctaagcaca agtcctacct
     4021 gaccccaggc ctgatcggct ctgtgggcac actggaggag ggcatccagg aggagcggaa
     4081 cgtgaaggag gacggagcac actctgccgc aaccgccaga tccccccagc cttgctctct
     4141 gctgggcaca aagttcaatt gtaaggagag ccccagaatc aacctgagga aggtgcgcaa
     4201 ggaggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4261 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4321 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4381 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4441 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4501 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4561 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4621 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4681 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4741 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4801 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4861 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4921 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4981 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5041 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5101 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5161 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5221 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5281 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5341 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ctggcctttt gctcacatgt t
//
