LOCUS       sequence-261799-        5829 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 5829)
  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 5829)
  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_SLC4A8
FEATURES             Location/Qualifiers
     source          1..5829
                     /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(3954..4053)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4070..4089)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4071..4089)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4094..4110)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4094..4110)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4212..4231)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4223..5032
                     /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(4290..4309)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5179..5767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5668..5687
                     /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 accatgcctg cagcaggatc taacgagcca gacggcgtgc tgagctacca
      721 gaggccagac gaggaggcag tggtggatca gggaggcacc tctacaatcc tgaatatcca
      781 ctacgagaag gaggagctgg agggccaccg gacactgtat gtgggcgtgc gcatgccact
      841 gggccggcag agccacagac accacaggac ccacggacag aagcaccgga gaaggggaag
      901 gggcaaggga gcaagccagg gagaggaggg cctggaggcc ctggcacacg acacaccttc
      961 ccagcgggtg cagtttatcc tgggcaccga ggaggatgag gagcacgtgc cacacgagct
     1021 gttcacagag ctggacgaga tctgcatgaa ggagggcgag gatgcagagt ggaaggagac
     1081 cgcaaggtgg ctgaagtttg aggaggacgt ggaggatgga ggagagcggt ggtctaagcc
     1141 ctacgtggcc acactgtccc tgcactctct gttcgagctg cggagctgtc tgatcaacgg
     1201 caccgtgctg ctggacatgc acgccaatag catcgaggag atctccgacc tgatcctgga
     1261 tcagcaggag ctgagctccg acctgaacga tagcatgaga gtgaaggtga gggaggccct
     1321 gctgaagaag caccaccacc agaatgagaa gaagcggaac aatctgatcc ctatcgtgag
     1381 gtccttcgcc gaagtgggca agaagcagtc tgacccacac ctgatggata agcacggaca
     1441 gaccgtgagc ccacagtccg tgcctaccac aaacctggag gtgaagaacg gcgtgaattg
     1501 cgagcactct cccgtggacc tgagcaaggt ggatctgcac tttatgaaga agatccctac
     1561 cggagcagag gcaagcaatg tgctggtggg cgaggtggac atcctggatc gcccaatcgt
     1621 ggccttcgtg cggctgtctc cagccgtgct gctgagcgga ctgacagagg tgcctatccc
     1681 aaccagattc ctgtttatcc tgctgggacc agtgggcaag ggacagcagt accacgagat
     1741 cggcaggtcc atggccacca tcatgacaga cgagatcttt cacgatgtgg cctataaggc
     1801 caaggagaga gacgatctgc tggccggcat cgacgagttc ctggatcagg tgaccgtgct
     1861 gccacctgga gagtgggacc caagcatccg catcgagcca cccaagaacg tgccctccca
     1921 ggagaagcgg aagatgcccg gcgtgcctaa cggcaacgtg tgccacatcg agcaggagcc
     1981 acacggagga cactccggac ctgagctgca gaggaccggc cggctgtttg gaggcctggt
     2041 gctggatatc aagcgcaagg ccccctggta ctggagcgac tatcgggatg ccctgtccct
     2101 gcagtgtctg gcctctttcc tgtttctgta ctgcgcctgt atgtcccctg tgatcacatt
     2161 cggcggactg ctgggagagg caaccgaggg aaggatctcc gccatcgagt ctctgtttgg
     2221 cgcctccatg accggcatcg cctattctct gttcgcagga caggccctga caatcctggg
     2281 aagcaccgga cccgtgctgg tgtttgagaa gatcctgttc aagttttgca aggactacgc
     2341 cctgtcttat ctgagcctga gagcctgcat cggcctgtgg acagccttcc tgtgcatcgt
     2401 gctggtggcc accgatgcct ctagcctggt gtgctacatc accaggttca cagaggaggc
     2461 ctttgcctcc ctgatctgta tcatctttat ctacgaggcc atcgagaagc tgatccacct
     2521 ggccgagacc tatcctatcc acatgcacag ccagctggac cacctgtccc tgtactattg
     2581 cagatgtaca ctgccagaga atcccaacaa tcacaccctg cagtattgga aggatcacaa
     2641 catcgtgaca gccgaggtgc actgggccaa tctgaccgtg tctgagtgcc aggagatgca
     2701 cggcgagttc atgggaagcg cctgtggaca ccacggacct tacacaccag acgtgctgtt
     2761 ttggtcctgc atcctgttct ttaccacatt catcctgtcc tctaccctga agacattcaa
     2821 gacctctagg tattttccaa cccgcgtgcg gtccatggtg tctgacttcg ccgtgtttct
     2881 gacaatcttc accatggtca tcatcgattt tctgatcgga gtgccaagcc ctaagctgca
     2941 ggtgccttcc gtgtttaagc caacaagaga cgataggggc tggatcatca acccaatcgg
     3001 cccaaatccc tggtggaccg tgatcgcagc aatcatcccc gccctgctgt gcacaatcct
     3061 gatcttcatg gaccagcaga tcaccgccgt gatcatcaac cgcaaggagc acaagctgaa
     3121 gaagggctgc ggctaccacc tggatctgct gatggtggcc atcatgctgg gcgtgtgcag
     3181 catcatgggc ctgccttggt tcgtggccgc cacagtgctg tctatcaccc acgtgaatag
     3241 cctgaagctg gagagcgagt gctccgcccc aggagagcag cccaagtttc tgggaatcag
     3301 ggagcagagg gtgacaggac tgatgatctt cgtgctgatg ggctgttccg tgttcatgac
     3361 cgccatcctg aagtttatcc ctatgccagt gctgtacggc gtgtttctgt atatgggcgt
     3421 gagctccctg cagggcatcc agttctttga ccggctgaag ctgtttggca tgcccgccaa
     3481 gcaccagcct gatttcatct atctgagaca cgtgcctctg aggaaggtgc acctgttcac
     3541 actgatccag ctgacctgcc tggtgctgct gtgggtcatc aaggcctccc cagccgccat
     3601 cgtgttcccc atgatggtgc tggccctggt gtttgtgaga aaagtgatgg acctgtgctt
     3661 ctccaagagg gagctgtctt ggctggacga tctgatgcca gagagcaaga agaagaagct
     3721 ggacgatgcc aagaagaagg ccaaggagga ggaggaggcc gagaagatgc tggagatcgg
     3781 cggcgataag tttcctctgg agtccagaaa gctgctgtct agcccaggca agaacatctc
     3841 ttgcaggtgt gaccccagcg agatcaatat ctccgatgag atgcctaaga ccacagtgtg
     3901 gaaggccctg tctatgaaca gcggcaatgc caaggagaag agcctgttca acgacccagc
     3961 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     4021 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     4081 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     4141 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     4201 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     4261 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     4321 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4381 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4441 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4501 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     4561 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4621 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     4681 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4741 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     4801 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4861 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4921 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4981 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     5041 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     5101 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     5161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5821 tcacatgtt
//
