LOCUS       sequence-259332-        3594 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 3594)
  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 3594)
  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_SLC10A2
FEATURES             Location/Qualifiers
     source          1..3594
                     /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(1719..1818)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1835..1854)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1836..1854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1859..1875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1977..1996)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1988..2797
                     /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(2055..2074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2944..3532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3433..3452
                     /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 accatgaacg atcccaattc ttgcgtggac aacgcaaccg tgtgcagcgg
      721 agcatcctgc gtggtgcctg agagcaattt caacaatatc ctgtccgtgg tgctgtctac
      781 cgtgctgaca atcctgctgg ccctggtcat gttctccatg ggctgtaacg tggagatcaa
      841 gaagtttctg ggccacatca agaggccttg gggcatctgc gtgggcttcc tgtgccagtt
      901 tggcatcatg ccactgaccg gcttcatcct gtctgtggcc tttgatatcc tgccactgca
      961 ggccgtggtg gtgctgatca tcggatgctg tccaggagga acagcaagca atatcctggc
     1021 ctactgggtg gacggcgata tggacctgtc tgtgagcatg accacatgct ccaccctgct
     1081 ggccctgggc atgatgccac tgtgcctgct gatctacaca aagatgtggg tggattccgg
     1141 ctctatcgtg atcccctatg acaacatcgg cacctctctg gtgagcctgg tggtgcccgt
     1201 gagcatcggc atgtttgtga atcacaagtg gcctcagaag gccaagatca tcctgaagat
     1261 cggctccatc gccggcgcca tcctgatcgt gctgatcgcc gtggtgggcg gcatcctgta
     1321 tcagtctgcc tggatcatcg ccccaaagct gtggatcatc ggcacaatct tccccgtggc
     1381 cggctactcc ctgggctttc tgctggcaag gatcgcagga ctgccttggt atcggtgcag
     1441 aaccgtggcc ttcgagacag gcatgcagaa cacccagctg tgcagcacaa tcgtgcagct
     1501 gtcctttacc cctgaggagc tgaatgtggt gttcacattt ccactgatct actctatctt
     1561 ccagctggcc tttgccgcca tcttcctggg cttttacgtg gcctataaga agtgccacgg
     1621 caagaacaag gccgagatcc ccgagagcaa ggagaatggc accgagcctg agagctcctt
     1681 ctataaggcc aacggcggct ttcagcccga cgagaaggac ccagctttct tgtacaaagt
     1741 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1801 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1861 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1921 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1981 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2041 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2101 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2161 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2221 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2281 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2341 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2401 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2461 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2521 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2581 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2641 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2701 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2761 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2821 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2881 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
