LOCUS       sequence-297829-        4599 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 4599)
  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 4599)
  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_SV2B
FEATURES             Location/Qualifiers
     source          1..4599
                     /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(2724..2823)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2840..2859)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2841..2859)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2864..2880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2864..2880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2982..3001)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2993..3802
                     /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(3060..3079)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3949..4537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4438..4457
                     /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 accatggacg attacaagta tcaggacaac tacggcggct atgccccttc
      721 cgatggctac tataggggca acgagtccaa tccagaggag gacgcccagt ctgatgtgac
      781 cgagggccac gacgaggagg atgagatcta cgagggcgag tatcagggca tcccccaccc
      841 tgacgatgtg aaggccaagc aggccaagat ggccccaagc cggatggact ccctgagagg
      901 ccagaccgat ctgatggcag agcggctgga ggacgaggag cagctggccc accagtacga
      961 gacaatcatg gatgagtgcg gccacggcag attccagtgg atcctgttct ttgtgctggg
     1021 actggccctg atggcagacg gagtggaggt gtttgtggtg agcttcgccc tgccatccgc
     1081 cgagaaggat atgtgcctga gctcctctaa gaagggcatg ctgggcatga tcgtgtatct
     1141 gggcatgatg gcaggagcct tcatcctggg aggactggca gacaagctgg gacggaagag
     1201 agtgctgtcc atgtctctgg ccgtgaacgc ctcctttgcc tctctgagct ccttcgtgca
     1261 gggctacggc gccttcctgt tttgccggct gatctccgga atcggaatcg gaggcgccct
     1321 gcccatcgtg ttcgcctatt tttctgagtt cctgagcagg gagaagaggg gagagcacct
     1381 gagctggctg ggcatcttct ggatgaccgg aggactgtac gcatctgcca tggcctggag
     1441 catcatccct cactacggct ggggctttag catgggcacc aattatcact ttcactcctg
     1501 gagagtgttc gtgatcgtgt gcgccctgcc atgtacagtg tctatggtgg ccctgaagtt
     1561 tatgccagag agccccaggt tcctgctgga gatgggcaag cacgacgagg cctggatgat
     1621 cctgaagcag gtgcacgata ccaacatgcg cgccaagggc acacccgaga aggtgttcac
     1681 cgtgagcaat atcaagacac ctaagcagat ggacgagttt atcgagatcc agtctagcac
     1741 cggcacatgg taccagaggt ggctggtgcg ctttaagaca atcttcaagc aagtgtggga
     1801 taacgccctg tactgcgtga tgggccccta tcggatgaat accctgatcc tggccgtggt
     1861 gtggtttgcc atggccttct cctactatgg cctgacagtg tggtttcctg acatgatcag
     1921 atacttccag gatgaggagt ataagtctaa gatgaaggtg ttctttggcg agcacgtgta
     1981 cggcgccaca atcaacttca ccatggagaa tcagatccac cagcacggca agctggtgaa
     2041 cgacaagttt accaggatgt atttcaagca cgtgctgttt gaggacacat tctttgatga
     2101 gtgctacttt gaggacgtga cctccacaga tacctatttc aagaactgta caatcgagtc
     2161 taccatcttt tacaataccg acctgtatga gcacaagttc atcaactgcc gctttatcaa
     2221 ttccacattc ctggagcaga aggagggctg tcacatggat ctggagcagg acaacgattt
     2281 tctgatctac ctggtgagct tcctgggcag cctgtccgtg ctgcctggca atatcatctc
     2341 cgccctgctg atggaccgga tcggcagact gaagatgatc ggcggctcca tgctgatctc
     2401 tgccgtgtgc tgtttctttc tgttctttgg caactctgag agcgccatga tcggctggca
     2461 gtgcctgttc tgtggcacca gcatcgccgc ctggaatgcc ctggatgtga tcacagtgga
     2521 gctgtatcca acaaaccagc gggccaccgc ctttggcatc ctgaatggcc tgtgcaagtt
     2581 cggcgccatc ctgggcaata ccatctttgc ctctttcgtg ggcatcacaa aggtggtgcc
     2641 aatcctgctg gcagcagcaa gcctggtggg aggaggcctg atcgccctga ggctgcccga
     2701 gacccgcgag caggtgctga tggacccagc tttcttgtac aaagttggca ttataagaaa
     2761 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2821 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2881 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2941 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3001 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3061 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3121 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3181 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3241 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3301 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3361 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3421 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3481 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3541 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3601 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3661 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3721 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3781 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     3841 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     3901 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4561 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
