LOCUS       sequence-263281-        5043 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 5043)
  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 5043)
  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_SLC39A10
FEATURES             Location/Qualifiers
     source          1..5043
                     /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(3168..3267)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3284..3303)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3285..3303)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3308..3324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3308..3324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3426..3445)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3437..4246
                     /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(3504..3523)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4393..4981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4882..4901
                     /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 accatgaagg tgcacatgca caccaagttc tgcctgatct gtctgctgac
      721 attcatcttt caccactgca accactgtca cgaggagcac gatcacggac cagaggccct
      781 gcacaggcag cacagaggca tgaccgagct ggagccctct aagttcagca agcaggccgc
      841 cgagaacgag aagaagtact atatcgagaa gctgtttgag cggtacggcg agaatggcag
      901 actgagcttc tttggcctgg agaagctgct gaccaacctg ggcctgggcg agaggaaggt
      961 ggtggagatc aatcacgagg atctgggcca cgaccacgtg tcccacctgg acatcctggc
     1021 cgtgcaggag ggcaagcact ttcactccca caaccaccag cactctcaca accacctgaa
     1081 tagcgagaac cagaccgtga caagcgtgtc caccaagcgc aatcacaagt gcgatcccga
     1141 gaaggagaca gtggaggtgt ccgtgaagtc tgacgataag cacatgcacg accacaacca
     1201 ccggctgaga caccaccaca ggctgcacca ccacctggat cacaacaata cccaccactt
     1261 ccacaatgac tctatcacac ctagcgagcg cggcgagcct tccaatgagc catctaccga
     1321 gacaaacaag acccaggagc agtccgatgt gaagctgcca aagggcaagc ggaagaagaa
     1381 gggccgcaag agcaacgaga attccgaagt gatcacaccc ggctttcccc ctaatcacga
     1441 tcagggcgag cagtatgagc acaacagggt gcacaagccc gaccgcgtgc acaatcctgg
     1501 acactcccac gtgcacctgc cagagaggaa cggacacgac ccaggaagag gacaccagga
     1561 cctggatcct gacaatgagg gcgagctgag gcacaccagg aagagggagg caccacacgt
     1621 gaagaacaat gccatcatca gcctgagaaa ggatctgaac gaggacgatc accaccacga
     1681 gtgtctgaat gtgacacagc tgctgaagta ctatggacac ggagcaaaca gcccaatctc
     1741 caccgacctg ttcacatacc tgtgccctgc cctgctgtat cagatcgatt cccggctgtg
     1801 catcgagcac tttgacaagc tgctggtgga ggatatcaat aaggacaaga acctggtgcc
     1861 agaggacgag gcaaacatcg gagcatctgc ctggatctgc ggcatcatca gcatcaccgt
     1921 gatctctctg ctgagcctgc tgggcgtgat cctggtgcct atcatcaatc agggctgttt
     1981 caagtttctg ctgaccttcc tggtggccct ggcagtggga acaatgagcg gcgatgcact
     2041 gctgcacctg ctgccacact cccagggagg acatgaccac tctcaccagc acgcacacgg
     2101 acacggacac tctcacggac acgagagcaa caagttcctg gaggagtacg atgccgtgct
     2161 gaagggactg gtggccctgg gaggcatcta tctgctgttc atcatcgagc actgcatcag
     2221 gatgtttaag cactacaagc agcagcgcgg caagcagaag tggtttatga agcagaatac
     2281 cgaggagagc acaatcggcc ggaagctgtc cgaccacaag ctgaacaata cccctgattc
     2341 tgactggctg cagctgaagc cactggccgg cacagacgat tccgtggtgt ctgaggatag
     2401 actgaacgag accgagctga cagacctgga gggacagcag gagagcccac caaagaatta
     2461 cctgtgcatc gaggaggaga agatcatcga tcactctcac agcgacggcc tgcacaccat
     2521 ccacgagcac gatctgcacg cagcagcaca caatcaccac ggagagaaca agacagtgct
     2581 gagaaagcac aaccaccagt ggcaccacaa gcacagccac cactcccacg gaccatgcca
     2641 ctccggatct gacctgaagg agaccggcat cgccaacatc gcctggatgg tcatcatggg
     2701 cgatggcatc cacaatttct ccgacggact ggcaatcgga gcagcctttt ctgccggact
     2761 gaccggagga atcagcacat ccatcgccgt gttctgtcac gagctgcctc acgagctggg
     2821 cgattttgcc gtgctgctga aggccggcat gaccgtgaag caggccatcg tgtacaatct
     2881 gctgagcgcc atgatggcct atatcggcat gctgatcggc acagccgtgg gccagtacgc
     2941 caacaatatc accctgtgga tcttcgccgt gacagccggc atgtttctgt atgtggccct
     3001 ggtggacatg ctgcccgaga tgctgcacgg cgatggcgac aacgaggagc acggcttctg
     3061 ccctgtgggc cagtttatcc tgcagaatct gggcctgctg ttcggctttg ccatcatgct
     3121 ggtcatcgcc ctgtacgagg ataagatcgt gttcgacatc cagtttgacc cagctttctt
     3181 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3241 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3301 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3361 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     3421 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     3481 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     3541 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     3601 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     3661 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     3721 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     3781 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     3841 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     3901 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     3961 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     4021 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     4081 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     4141 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     4201 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     4261 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     4321 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     4381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4501 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5041 gtt
//
