LOCUS       sequence-261080-        3930 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 3930)
  AUTHORS   Polesel M, Ingles-Prieto A, Christodoulaki E, Ferrada E, Doucerain 
            C, Altermatt P, Knecht M, Kuhn M, Steck AL, Wilhelm M, Manolova V
  TITLE     Functional characterization of SLC39 family members ZIP5 and ZIP10 
            in overexpressing HEK293 cells reveals selective copper transport 
            activity.
  JOURNAL   Biometals. 2022 Dec 1. doi: 10.1007/s10534-022-00474-6.
  PUBMED    36454509
REFERENCE   2  (bases 1 to 3930)
  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"; journalName: "Biometals. 
            2022 Dec 1. doi: 10.1007/s10534-022-00474-6."
COMMENT     Sequence Label: pDONR221_SLC39A8
FEATURES             Location/Qualifiers
     source          1..3930
                     /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(2055..2154)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2171..2190)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2172..2190)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2195..2211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2195..2211)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2313..2332)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2324..3133
                     /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(2391..2410)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3280..3868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3769..3788
                     /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 accatggccc ctggccgggc cgtggccggc ctgctgctgc tggcagcagc
      721 aggactggga ggagtggccg agggcccagg cctggccttt tccgaggacg tgctgagcgt
      781 gttcggagca aacctgtccc tgtctgccgc acagctgcag cacctgctgg agcagatggg
      841 agcagcatcc agagtgggag tgccagagcc tggccagctg cacttcaacc agtgtctgac
      901 cgccgaggag atctttagcc tgcacggctt ctccaatgcc acccagatca caagctccaa
      961 gttttctgtg atctgccccg ccgtgctgca gcagctgaac ttccacccgt gcgaggatag
     1021 gccgaaacat aagacaagac ctagccactc cgaagtgtgg ggctacggct ttctgtctgt
     1081 gaccatcatc aatctggcca gcctgctggg cctgatcctg acaccactga tcaagaagtc
     1141 ctatttcccc aagatcctga ccttctttgt gggcctggcc atcggcacac tgttttctaa
     1201 cgccatcttc cagctgatcc ccgaggcctt tggcttcgac cctaaggtgg atagctacgt
     1261 ggagaaggcc gtggccgtgt ttggcggctt ctatctgctg ttctttttcg agaggatgct
     1321 gaagatgctg ctgaagacct acggccagaa tggccacaca cactttggca acgacaattt
     1381 cggcccccag gagaagaccc accagccaaa ggccctgcca gccatcaacg gagtgacctg
     1441 ctatgccaat cctgccgtga cagaggccaa cggccacatc cactttgaca acgtgagcgt
     1501 ggtgtctctg caggatggca agaaggagcc ttctagctgc acctgtctga agggcccaaa
     1561 gctgtccgag atcggcacca tcgcctggat gatcacactg tgcgacgccc tgcacaactt
     1621 catcgatggc ctggccatcg gcgcctcttg taccctgagc ctgctgcagg gcctgtctac
     1681 aagcatcgcc atcctgtgcg aggagtttcc ccacgagctg ggcgacttcg tgatcctgct
     1741 gaacgccggc atgtccacca gacaggccct gctgtttaat ttcctgagcg cctgctcctg
     1801 ttacgtggga ctggccttcg gcatcctggt gggcaacaat tttgccccaa atatcatctt
     1861 cgccctggcc ggcggcatgt ttctgtatat cagcctggcc gacatgttcc ccgagatgaa
     1921 cgatatgctg agggagaagg tgaccggccg caagaccgac ttcaccttct tcatgatcca
     1981 gaatgccggc atgctgaccg gcttcacagc catcctgctg atcacactgt acgccggcga
     2041 gatcgagctg gaggacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2101 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2161 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2221 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2281 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2341 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2401 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2461 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2521 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2581 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2641 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2701 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2761 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2821 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2881 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2941 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3001 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3061 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3121 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3181 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3421 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3481 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3541 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3601 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3661 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3721 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3781 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3841 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3901 ggccttttgc tggccttttg ctcacatgtt
//
