LOCUS       sequence-262927-        4170 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 4170)
  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 4170)
  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_SLC39A5
FEATURES             Location/Qualifiers
     source          1..4170
                     /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(2295..2394)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2411..2430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2412..2430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2435..2451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2435..2451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2553..2572)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2564..3373
                     /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(2631..2650)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3520..4108
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4009..4028
                     /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 accatgatgg gatctcccgt gagccacctg ctggcaggat tctgcgtgtg
      721 ggtggtgctg ggatgggtgg gaggatccgt gccaaatctg ggacctgcag agcaggagca
      781 gaaccactac ctggcccagc tgtttggcct gtatggagag aatggcaccc tgacagcagg
      841 aggactggca aggctgctgc actctctggg cctgggccgg gtgcagggcc tgagactggg
      901 ccagcacgga ccactgaccg gaagggcagc atcccctgcc gccgataatt ctacacaccg
      961 cccccagaac cctgagctga gcgtggacgt gtgggcagga atgccactgg gaccaagcgg
     1021 ctggggcgat ctggaggagt ccaaggcacc tcacctgcca aggggaccag cacccagcgg
     1081 cctggacctg ctgcacagac tgctgctgct ggaccactcc ctggccgatc acctgaacga
     1141 ggactgcctg aatggctccc agctgctggt gaacttcgga ctgtctccag cagcacctct
     1201 gaccccaagg cagtttgccc tgctgtgccc tgccctgctg taccagatcg acagccgggt
     1261 gtgcatcgga gcacctgcac cagcaccacc tggcgatctg ctgtctgccc tgctgcagag
     1321 cgccctggcc gtgctgctgc tgtccctgcc atctcctctg agcctgctgc tgctgaggct
     1381 gctgggacct aggctgctga ggccactgct gggcttcctg ggcgccctgg cagtgggcac
     1441 cctgtgcggc gacgccctgc tgcacctgct gcctcacgca caggagggca gacacgcagg
     1501 accaggagga ctgcccgaga aggatctggg accaggactg tccgtgctgg gaggactgtt
     1561 cctgctgttt gtgctggaga atatgctggg actgctgagg cacaggggac tgaggcccag
     1621 atgctgtcgg agaaagaggc gcaacctgga gacccggaat ctggacccag agaacggctc
     1681 tggcatggcc ctgcagccac tgcaggcagc accagagcca ggagcacagg gacagagaga
     1741 gaagaattct cagcacccac ccgccctggc acctccagga caccagggac acagccacgg
     1801 ccaccagggc ggcaccgata tcacatggat ggtgctgctg ggcgatggcc tgcacaacct
     1861 gacagacgga ctggcaatcg gagcagcctt ctctgacggc tttagctccg gcctgagcac
     1921 cacactggcc gtgttttgtc acgagctgcc acacgagctg ggcgatttcg ccatgctgct
     1981 gcagtccggc ctgtcttttc ggagactgct gctgctgagc ctggtgtccg gcgccctggg
     2041 actgggagga gccgtgctgg gcgtgggact gtccctggga cctgtgccac tgaccccatg
     2101 ggtgttcgga gtgacagcag gcgtgtttct gtatgtggcc ctggtggata tgctgcctgc
     2161 cctgctgagg ccacctgagc ccctgcctac cccacacgtg ctgctgcagg gactgggact
     2221 gctgctggga ggaggactga tgctggcaat cacactgctg gaggagcgcc tgctgccagt
     2281 gaccacagag ggcgacccag ctttcttgta caaagttggc attataagaa agcattgctt
     2341 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2401 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2461 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2521 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2581 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2641 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2701 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2761 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2821 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2881 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2941 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     3001 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     3061 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     3121 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     3181 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     3241 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     3301 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     3361 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     3421 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3481 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3541 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3601 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3661 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3721 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3781 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3841 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3901 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3961 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4021 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4081 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4141 ggccttttgc tggccttttg ctcacatgtt
//
