LOCUS       sequence-261770-        4224 bp    DNA     circular SYN 14-APR-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 4224)
  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 4224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_SLC22A31
FEATURES             Location/Qualifiers
     source          1..4224
                     /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(2349..2448)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2465..2484)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2466..2484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2489..2505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2489..2505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2607..2626)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2618..3427
                     /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(2685..2704)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3574..4162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4063..4082
                     /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 accatggagc aggaggcacg cgtgctgagg gcagcaggag gcttcggaag
      721 agcacggaga ctgctggcat ccgcctcttg ggtgccatgc atcgtgctgg gcctggtgct
      781 gagctccgag gagctgctga ccgcacagcc agcacctcac tgtaggccag accccacact
      841 gctgccacct gccctgagag ccctgagggg acccgccctg ctggatgcag caatcccaag
      901 actgggacct accagggcac cagcagaggc cctgggcgtg ctgagcccat cctacctggc
      961 acctctgaca agggcaccta ggccatctag ctgggcatct tgcagcggag cagcagcagg
     1021 acctacctgg aacctggtgt gcggcgacgg atggaaggtg ccactggagc aggtgagcca
     1081 cctgctggga tggctgctgg gatgcgtgat cctgggagca ggatgtgata ggttcggcag
     1141 gcgcgccgtg tttgtggcat ccctggtgct gaccacagga ctgggagcat ctgaggccct
     1201 ggcagcaagc ttccccaccc tgctggtgct gaggctgctg cacggaggca cactggcagg
     1261 cgccctgctg gccctgtatc tggccagact ggagctgtgc gacccaccac acaggctggc
     1321 cttctccatg ggagcaggcc tgttttctgt ggtgggcacc ctgctgctgc ctggactggc
     1381 cgccctggtg caggattgga gactgctgca gggactgggc gccctgatgt ccggcctgct
     1441 gctgctgttc tggggctttc cagccctgtt cccagagtct ccctgttggc tgctggcaac
     1501 aggacaggtg gcaagggcaa ggaagatcct gtggcggttt gcagaggcat ctggagtggg
     1561 accaggcgac tcctctctgg aggagaacag cctggccacc gccacagagc tgaccatgct
     1621 gtctgcccgc agcccacagc ctaggtacca cagccctctg ggactgctga ggaccagagt
     1681 gacatggcgc aatggcctga tcctgggctt cagctccctg gtgggaggag gaatcagggc
     1741 ctcctttcgg agatctctgg cccctcaggt gccaacattc tacctgcctt attttctgga
     1801 ggcaggactg gaggcagccg ccctggtgtt cctgctgctg accgcagatt gctgtggcag
     1861 gcgcccagtg ctgctgctgg gaaccatggt gacaggactg gcaagcctgc tgctgctggc
     1921 aggagcacag tatctgccag gctggacagt gctgtttctg tccgtgctgg gactgctggc
     1981 aagcagagcc gtgtccgccc tgtctagcct gttcgcagca gaggtgtttc ccacagtgat
     2041 caggggagca ggactgggac tggtgctggg agccggcttc ctgggccagg ccgccggccc
     2101 tctggacacc ctgcacggcc gccagggctt ctttctgcag caggtggtgt ttgcatccct
     2161 ggccgtgctg gccctgctgt gcgtgctgct gctgccagag tccaggtcta ggggactgcc
     2221 acagagcctg caggacgcag ataggctgcg gagatcccca ctgctgaggg gcagacccag
     2281 gcaggatcac ctgcctctgc tgcctccaag caattcctac tgggccggcc acacccccga
     2341 gcagcacgac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2401 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2461 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2521 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
     2581 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     2641 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     2701 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
     2761 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     2821 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     2881 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
     2941 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     3001 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     3061 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     3121 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     3181 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     3241 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     3301 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     3361 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     3421 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     3481 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3721 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4201 ttgctggcct tttgctcaca tgtt
//
