LOCUS       sequence-478921-        5832 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Gateway entry clone with codon-optimized ORF sequence for subcloning
            into custom expression vectors. Contains a STOP codon at the end of 
            the codon-optimized ORF sequence. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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_STOP
FEATURES             Location/Qualifiers
     source          1..5832
                     /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(3957..4056)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4073..4092)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4074..4092)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4097..4113)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4097..4113)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4215..4234)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4226..5035
                     /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(4293..4312)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5182..5770
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5671..5690
                     /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 actaagaccc
     3961 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     4021 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     4081 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     4141 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     4201 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     4261 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     4321 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     4381 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     4441 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     4501 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     4561 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     4621 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     4681 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     4741 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     4801 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     4861 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     4921 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     4981 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     5041 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     5101 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     5161 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5221 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5281 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5341 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5401 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5461 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5521 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5581 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5641 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5701 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5761 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5821 tgctcacatg tt
//
