LOCUS       sequence-260456-        5469 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 5469)
  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 5469)
  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_SLC8A1
FEATURES             Location/Qualifiers
     source          1..5469
                     /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(3594..3693)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3710..3729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3711..3729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3734..3750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3734..3750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3852..3871)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3863..4672
                     /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(3930..3949)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4819..5407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5308..5327
                     /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 accatgtaca acatgcggag actgtccctg tctcctacct tcagcatggg
      721 ctttcacctg ctggtgacag tgagcctgct gttctcccac gtggatcacg tgatcgccga
      781 gaccgagatg gagggcgagg gcaatgagac aggcgagtgc accggctcct actattgtaa
      841 gaagggcgtg atcctgccaa tctgggagcc acaggatccc tcttttggcg acaagatcgc
      901 cagagccaca gtgtatttcg tggccatggt gtacatgttt ctgggcgtgt ctatcatcgc
      961 cgacaggttc atgagctcca tcgaagtgat cacaagccag gagaaggaga tcaccatcaa
     1021 gaagccaaac ggcgagacca caaagaccac agtgagaatc tggaacgaga cagtgagcaa
     1081 tctgaccctg atggcactgg gatctagcgc cccagagatc ctgctgtccg tgatcgaggt
     1141 gtgcggccac aactttacag ccggcgatct gggaccttct accatcgtgg gaagcgccgc
     1201 cttcaatatg tttatcatca tcgccctgtg cgtgtacgtg gtgcctgacg gcgagaccag
     1261 aaagatcaag cacctgaggg tgttctttgt gaccgccgcc tggagcatct tcgcctatac
     1321 atggctgtac atcatcctgt ccgtgatctc tccaggagtg gtggaagtgt gggagggact
     1381 gctgacattc tttttctttc ccatctgcgt ggtgttcgca tgggtggcag acaggcgcct
     1441 gctgttttac aagtacgtgt acaagagata cagggccggc aagcagagag gcatgatcat
     1501 cgagcacgag ggcgataggc cctcctctaa gaccgagatc gagatggacg gcaaggtggt
     1561 gaactcccac gtggagaatt ttctggatgg cgccctggtg ctggaggtgg atgagaggga
     1621 ccaggacgat gaggaggcaa ggagagagat ggcccggatc ctgaaggagc tgaagcagaa
     1681 gcaccctgac aaggagatcg agcagctgat cgagctggcc aactatcagg tgctgtccca
     1741 gcagcagaag tctcgcgcct tctaccggat ccaggcaaca aggctgatga ccggagcagg
     1801 caatatcctg aagaggcacg cagcagatca ggcaaggaag gccgtgtcca tgcacgaggt
     1861 gaacacagag gtgaccgaga atgaccccgt gtctaagatc ttctttgagc agggcaccta
     1921 tcagtgcctg gagaactgtg gcacagtggc cctgaccatc atcaggaggg gaggcgatct
     1981 gaccaacaca gtgttcgtgg attttcgcac agaggacgga accgcaaatg caggatccga
     2041 ttacgagttc acagagggca ccgtggtgtt taagcctggc gacacccaga aggagatccg
     2101 ggtgggcatc atcgacgatg acatcttcga ggaggatgag aactttctgg tgcacctgtc
     2161 taatgtgaag gtgagctccg aggcaagcga ggacggaatc ctggaggcca accacgtgtc
     2221 caccctggca tgcctgggaa gcccatccac cgccacagtg accatcttcg atgacgatca
     2281 cgccggcatc ttcacatttg aggagcccgt gacccacgtg tctgagagca tcggcatcat
     2341 ggaggtgaag gtgctgagaa cctctggcgc caggggcaat gtgatcgtgc catataagac
     2401 aatcgaggga accgcaaggg gaggaggaga ggatttcgag gacacctgtg gcgagctgga
     2461 gtttcagaac gacgagatcg tgaagacaat cagcgtgaaa gtgatcgacg atgaggagta
     2521 cgagaagaat aagaccttct ttctggagat cggagagcca aggctggtgg agatgtccga
     2581 gaagaaggcc ctgctgctga atgagctggg cggcttcaca atcaccggca agtatctgtt
     2641 cggccagccc gtgtttcgca aggtgcacgc ccgggagcac cctatcctgt ctacagtgat
     2701 caccatcgcc gatgagtacg acgataagca gcctctgacc agcaaggagg aggaggagcg
     2761 gagaatcgcc gagatgggca gaccaatcct gggcgagcac acaaagctgg aagtgatcat
     2821 cgaggagagc tatgagttca agtccacagt ggacaagctg atcaagaaga ccaacctggc
     2881 cctggtggtg ggcacaaata gctggaggga gcagtttatc gaggcaatca ccgtgtccgc
     2941 cggagaggac gatgacgatg acgagtgcgg cgaggagaag ctgccctctt gtttcgatta
     3001 tgtgatgcac ttcctgaccg tgttttggaa ggtgctgttc gcctttgtgc cccctacaga
     3061 gtactggaac ggctgggcct gcttcatcgt gagcatcctg atgatcggcc tgctgaccgc
     3121 cttcatcggc gatctggcct cccactttgg ctgtaccatc ggcctgaagg actctgtgac
     3181 agccgtggtg ttcgtggccc tgggcacatc tgtgcctgac acctttgcca gcaaggtggc
     3241 cgccacccag gatcagtatg ccgacgcctc catcggcaat gtgacaggct ctaacgccgt
     3301 gaacgtgttc ctgggaatcg gagtggcatg gagcatcgca gcaatctacc acgcagcaaa
     3361 cggagagcag tttaaggtga gcccaggcac cctggccttc tccgtgacac tgtttaccat
     3421 cttcgccttt atcaatgtgg gcgtgctgct gtacaggcgc cggcctgaga tcggaggaga
     3481 gctgggagga ccaaggacag caaagctgct gacctcctgc ctgttcgtgc tgctgtggct
     3541 gctgtatatc ttcttttcta gcctggaggc ctactgtcac atcaagggct ttgacccagc
     3601 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     3661 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     3721 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     3781 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     3841 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     3901 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     3961 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     4021 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     4081 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     4141 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     4201 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     4261 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     4321 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     4381 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     4441 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     4501 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     4561 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     4621 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     4681 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     4741 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     4801 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4861 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4921 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     4981 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5041 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5101 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5161 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5221 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5281 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5341 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5401 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5461 tcacatgtt
//
