LOCUS       sequence-263888-        3714 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 3714)
  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 3714)
  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_SLC30A3
FEATURES             Location/Qualifiers
     source          1..3714
                     /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(1839..1938)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1955..1974)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1956..1974)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1979..1995)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1979..1995)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2097..2116)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2108..2917
                     /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(2175..2194)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3064..3652
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3553..3572
                     /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 cttccccagc agcaggagga ctggagacca caaggctggt
      721 gtctccaagg gacaggggag gagcaggagg atccctgcgc ctgaagtctc tgtttacaga
      781 gccctctgag cccctgcctg aggagagcaa gcctgtggag atgccattcc accactgcca
      841 cagggatcca ctgccacctc caggactgac ccctgagaga ctgcacgcac ggagacagct
      901 gtacgcagca tgcgccgtgt gcttcgtgtt tatggcagga gaggtggtgg gaggatatct
      961 ggcacactcc ctggccatca tgacagacgc agcacacctg ctggcagatg tgggatctat
     1021 gatgggctcc ctgttttctc tgtggctgag caccaggcca gcaaccagaa caatgacctt
     1081 cggctggcac aggtctgaga cactgggcgc cctggcaagc gtggtgtccc tgtggatggt
     1141 gaccggcatc ctgctgtacc tggcctttgt gcgcctgctg cacagcgact atcacatcga
     1201 gggaggagca atgctgctga cagcatccat cgccgtgtgc gccaacctgc tgatggcctt
     1261 cgtgctgcac caggcaggac cacctcactc tcacggaagc aggggagcag agtacgcacc
     1321 actggaggag ggccctgagg agccactgcc cctgggcaat accagcgtga gagcagcctt
     1381 cgtgcacgtg ctgggcgacc tgctgcagtc ctttggcgtg ctggccgcct ctatcctgat
     1441 ctacttcaag ccacagtata aggccgccga tcccatcagc acattcctgt tttccatctg
     1501 tgcactggga tctacagcac ccaccctgag ggacgtgctg aggatcctga tggagggcac
     1561 ccctaggaac gtgggctttg agccagtgcg cgatacactg ctgagcgtgc ctggcgtgcg
     1621 ggcaacccac gagctgcacc tgtgggccct gacactgacc tatcacgtgg caagcgccca
     1681 cctggcaatc gactccacag cagatcctga ggccgtgctg gcagaggcaa gctccaggct
     1741 gtactccaga ttcggctttt ctagctgtac cctgcaggtg gagcagtatc agccagagat
     1801 ggcacagtgc ctgaggtgtc aggagccacc ccaggccgac ccagctttct tgtacaaagt
     1861 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1921 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1981 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2041 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     2101 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2161 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2221 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2281 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2341 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2401 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2461 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2521 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2581 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2641 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2701 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2761 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2821 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2881 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2941 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     3001 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3061 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3121 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3181 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3241 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3301 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3361 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3421 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3481 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3541 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3601 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3661 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
