LOCUS       sequence-265777-        4623 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 4623)
  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 4623)
  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_SLC39A12
FEATURES             Location/Qualifiers
     source          1..4623
                     /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(2748..2847)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2864..2883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2865..2883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2888..2904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2888..2904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3006..3025)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3017..3826
                     /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(3084..3103)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3973..4561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4462..4481
                     /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 accatgtgct tccgcaccaa gctgagcgtg tcctgggtgc ctctgttcct
      721 gctgctgtct cgggtgttta gcacagagac cgacaagcca tctgcccagg attctcgcag
      781 caggggaagc tccggacagc ctgcagacct gctgcaggtg ctgagcgccg gcgatcaccc
      841 acctcacaac cactcccgct ctctgatcaa gaccctgctg gagaagacag gctgccctcg
      901 gagaaggaac ggcatgcagg gcgactgcaa cctgtgcttc gagccagatg cactgctgct
      961 gatcgcagga ggcaattttg aggaccagct gagggaggag gtggtgcagc gggtgtccct
     1021 gctgctgctg tactatatca tccaccagga ggagatctgt tctagcaagc tgaacatgtc
     1081 taataaggag tacaagttct atctgcactc tctgctgagc ctgagacagg acgaggattc
     1141 ctcttttctg agccagaacg agaccgagga catcctggcc ttcaccaggc agtactttga
     1201 tacaagccag tcccagtgca tggagacaaa gaccctgcag aagaagagcg gaatcgtgag
     1261 ctccgaggga gcaaacgagt ccaccctgcc acagctggcc gccatgatca tcacactgtc
     1321 cctgcagggc gtgtgcctgg gacagggaaa tctgccttct ccagactact tcaccgagta
     1381 tatcttttct agcctgaaca gaacaaatac cctgaggctg agcgagctgg atcagctgct
     1441 gaacacactg tggaccagat ccacatgcat caagaatgag aagatccacc agttccagag
     1501 gaagcagaac aatatcatca cccacgacca ggattacagc aatttttcct ctagcatgga
     1561 gaaggagtct gaggacggcc ccgtgagctg ggatcagaca tgtttctccg ccagacagct
     1621 ggtggagatc tttctgcaga agggcctgtc cctgatctct aaggaggact tcaagcagat
     1681 gagcccaggc atcatccagc agctgctgtc ttgcagctgt cacctgccca aggatcagca
     1741 ggccaagctg ccacccacca cactggagaa gtacggctat tctaccgtgg ccgtgaccct
     1801 gctgacactg ggcagcatgc tgggcacagc cctggtgctg ttccactcct gcgaggagaa
     1861 ctataggctg atcctgcagc tgtttgtggg actggcagtg ggcaccctga gcggcgacgc
     1921 actgctgcac ctgatccctc aggtgctggg actgcacaag caggaggcac cagagttcgg
     1981 ccactttcac gagtccaagg gccacatctg gaagctgatg ggcctgatcg gcggcatcca
     2041 cggcttcttt ctgatcgaga agtgtttcat cctgctggtg tcccccaacg ataagcaggg
     2101 cctgtctctg gtgaatggac acgtgggaca ctcccaccac ctggccctga atagcgagct
     2161 gtccgaccag gcaggaaggg gcaagtccgc ctctaccatc cagctgaagt cccccgagga
     2221 ttctcaggcc gccgagatgc ctatcggcag catgacagcc tccaaccgga agtgcaaggc
     2281 catctctctg ctggccatca tgatcctggt gggcgacagc ctgcacaatt tcgcagatgg
     2341 actggcaatc ggagcagcct tttcctctag ctccgagagc ggcgtgacca caaccatcgc
     2401 catcctgtgc cacgagatcc cccacgagat gggcgacttt gccgtgctgc tgtctagcgg
     2461 cctgtccatg aagaccgcca tcctgatgaa cttcatctcc tctctgacag cctttatggg
     2521 cctgtacatc ggactgagcg tgtccgccga cccttgcgtg caggattgga tcttcacagt
     2581 gaccgccggc atgtttctgt atctgagcct ggtggagatg ctgccagaga tgacccacgt
     2641 gcagacacag cggccctgga tgatgttcct gctgcagaac tttggcctga tcctgggatg
     2701 gctgtccctg ctgctgctgg ccatctatga gcagaatatc aagatcgacc cagctttctt
     2761 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     2821 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     2881 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     2941 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3001 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     3061 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3121 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3181 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3241 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3301 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3361 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3421 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3481 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3541 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     3601 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     3661 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     3721 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     3781 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     3841 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     3901 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     3961 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4021 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4081 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4141 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4201 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4261 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4321 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4381 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4441 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4501 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4561 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4621 gtt
//
