LOCUS       sequence-263266-        5961 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 5961)
  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 5961)
  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_SLC4A5
FEATURES             Location/Qualifiers
     source          1..5961
                     /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(4086..4185)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4202..4221)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4203..4221)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4226..4242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4226..4242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4344..4363)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4355..5164
                     /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(4422..4441)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5311..5899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5800..5819
                     /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 accatgaagg tgaaggagga gaaggccggc gtgggcaagc tggaccacac
      721 caaccaccgg agaaggttcc ccgatcagaa ggagtgccca cctatccaca tcggactgcc
      781 agtgcctacc taccctcaga gaaagacaga tcagaagggc cacctgtctg gcctgcagaa
      841 ggtgcactgg ggactgaggc cagaccagcc acagcaggag ctgaccggac ctggaagcgg
      901 agcaagctcc caggactcta gcatggatct gatctctaga acaaggagcc cagcagcaga
      961 gcagctgcag gacatcctgg gagaggagga tgaggcccca aatcccaccc tgtttacaga
     1021 gatggatacc ctgcagcacg acggcgatca gatggagtgg aaggagagcg cccgctggat
     1081 caagttcgag gagaaggtgg aggagggagg agagcggtgg tccaagcccc acgtgagcac
     1141 cctgtccctg cactctctgt tcgagctgag aacatgcctg cagaccggca cagtgctgct
     1201 ggacctggat tctggcagcc tgccacagat catcgacgat gtgatcgaga agcagatcga
     1261 ggacggactg ctgaggccag agctgaggga gcgggtgtct tacgtgctgc tgcgccggca
     1321 caggcaccag accaagaagc ctatccaccg gagcctggcc gatatcggca agtccgtgtc
     1381 taccacaaac agatcccctg caaggtctcc aggagcagga ccaagcctgc accactccac
     1441 agaggacctg agaatgaggc agtccgccaa ctatggccgg ctgtgccacg cacagagccg
     1501 gtccatgaat gacatctctc tgacccctaa cacagatcag cggaagaata agtttatgaa
     1561 gaagatccca aaggacagcg aggcctccaa tgtgctggtg ggcgaggtgg acttcctgga
     1621 tcagcccttc atcgcctttg tgagactgat ccagtccgcc atgctgggag gagtgaccga
     1681 ggtgcctgtg ccaacaaggt tcctgtttat cctgctgggc cctagcggca gagccaagtc
     1741 ctacaacgag atcggcaggg ccatcgccac cctgatggtg gacgatctgt ttagcgacgt
     1801 ggcctataag gcccgcaatc gggaggatct gatcgccggc atcgacgagt tcctggatga
     1861 agtgatcgtg ctgccaccag gagagtggga cccaaacatc agaatcgagc ctccaaagaa
     1921 ggtgcccagc gccgataaga ggaagagcgt gttcagcctg gccgagctgg gccagatgaa
     1981 cggctccgtg ggaggaggag gaggagcacc aggaggaggc aatggcggag gaggcggcgg
     2041 cggctctggc ggcggcgccg gcagcggcgg cgccggcggc acctcctctg gcgacgatgg
     2101 cgagatgccc gccatgcacg agatcggcga ggagctgatc tggacaggcc gcttctttgg
     2161 cggcctgtgc ctggatatca agcggaagct gccttggttc ccatccgact tttacgatgg
     2221 cttccacatc cagtctatca gcgccatcct gtttatctat ctgggctgta tcaccaacgc
     2281 catcacattc ggcggactgc tgggcgacgc aaccgataat taccagggcg tgatggagtc
     2341 ctttctgggc acagccatgg ccggctccct gttctgcctg ttttctggcc agcccctgat
     2401 catcctgagc tccaccggcc ctatcctgat ctttgagaag ctgctgttcg actttagcaa
     2461 gggcaacggc ctggattata tggagttccg cctgtggatc ggactgcact ccgccgtgca
     2521 gtgtctgatc ctggtggcca ccgacgcctc tttcatcatc aagtacatca cccggtttac
     2581 agaggagggc ttctctacac tgatcagctt catctttatc tatgacgcca tcaagaagat
     2641 gatcggcgcc tttaagtact atccaatcaa catggacttc aagcccaatt tcatcaccac
     2701 atacaagtgc gagtgcgtgg cccccgacac cgtgaacacc acagtgttca atgcaagcgc
     2761 cccactggca cctgacacca acgcctccct gtacaacctg ctgaatctga cagccctgga
     2821 ttggtccctg ctgtctaaga aggagtgcct gtcttatggc ggccgcctgc tgggcaatag
     2881 ctgtaagttt atccctgatc tggccctgat gtccttcatc ctgttctttg gcacctactc
     2941 tatgaccctg acactgaaga agttcaagtt tagcagatat ttcccaacaa aggtgagggc
     3001 cctggtggcc gacttttcca tcgtgttctc tatcctgatg ttttgcggca tcgatgcctg
     3061 tttcggactg gagaccccaa agctgcacgt gcctagcgtg atcaagccaa caagacccga
     3121 caggggctgg tttgtggccc cattcggcaa gaacccctgg tgggtgtatc ctgcctccat
     3181 cctgccagcc ctgctggtga ccatcctgat ctttatggat cagcagatca cagccgtgat
     3241 cgtgaacaga aaggagaata agctgaagaa ggccgccggc taccacctgg acctgttttg
     3301 ggtgggcatc ctgatggccc tgtgcagctt catgggcctg ccttggtacg tggccgccac
     3361 cgtgatcagc atcgcccaca tcgattccct gaagatggag accgagacat ccgcccctgg
     3421 agagcagcca cagttcctgg gcgtgcgcga gcagagggtg accggcatca tcgtgtttat
     3481 cctgacaggc atcagcgtgt tcctggcccc aatcctgaag tgtatccccc tgcctgtgct
     3541 gtacggcgtg tttctgtata tgggcgtggc ctctctgaat ggaatccaga tgggaaccgg
     3601 aggaagcgag ttcaagatcc agaagaagct gacacccttt tgggagcggt gcaagctgtt
     3661 cctgatgccc gccaagcacc agcctgacca cgcctttctg cggcacgtgc ctctgagaag
     3721 gatccacctg ttcaccctgg tgcagatcct gtgcctggcc gtgctgtgga tcctgaagtc
     3781 cacagtggcc gccatcatct ttcccgtgat gatcctgggc ctgatcatcg tgcgccggct
     3841 gctggatttc atcttttctc agcacgacct ggcctggatc gataacatcc tgccagagaa
     3901 ggagaagaag gagaccgaca agaagagaaa gaggaagaag ggagcacacg aggactgcga
     3961 tgaggagccc cagttccccc ctccaagcgt gatcaagatc cctatggaga gcgtgcagtc
     4021 cgacccacag aatggcatcc actgtatcgc ccgcaagcgg tctagctcct ggtcctactc
     4081 tctggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4141 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4201 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4261 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4321 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4381 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4441 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4501 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4561 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4621 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4681 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4741 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4801 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4861 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     4921 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     4981 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5041 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5101 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     5161 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     5221 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     5281 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5341 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5401 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5461 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5521 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5581 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5641 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5701 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5761 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5821 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5881 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5941 ctggcctttt gctcacatgt t
//
