LOCUS       sequence-150710-        5931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express COMMD8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5931)
  AUTHORS   Burstein E, Hoberg JE, Wilkinson AS, Rumble JM, Csomos RA, Komarck 
            CM, Maine GN, Wilkinson JC, Mayo MW, Duckett CS
  TITLE     COMMD proteins, a novel family of structural and functional homologs
            of MURR1.
  JOURNAL   J Biol Chem. 2005 Jun 10;280(23):22222-32. Epub 2005 Mar 30.
  PUBMED    15799966
REFERENCE   2  (bases 1 to 5931)
  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: "J Biol 
            Chem."; date: "2005-06-10"; volume: "280(23)"; pages: "22222-32. 
            Epub 2005 Mar 30"
COMMENT     Sequence Label: pEBB COMMD8 Flag
FEATURES             Location/Qualifiers
     source          1..5931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        252..462
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        274..470
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      321..456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        557..1739
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          788..1730
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1687..1707
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1788..1797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2352..2375
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3065..3082)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3065..3081)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3074..3096)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3294..3749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3591..3612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3766..3785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3885..3907
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3945..3963)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4031..4135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4136..4204,4205..4996)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4354..4373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5167..5755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5656..5675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5909..5926
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttc
      241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      301 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      361 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      421 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttgca
      481 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
      541 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
      601 ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg
      661 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
      721 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
      781 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
      841 ttgcgtgcct tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc
      901 ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc
      961 gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc
     1021 ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg
     1081 ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg
     1141 gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt
     1201 cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg
     1261 gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa
     1321 ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg
     1381 cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac
     1441 aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg
     1501 cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg
     1561 gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact gaagttaggc
     1621 cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt
     1681 tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag gtgtcgtgag
     1741 gaattctcta gagatccctc gacctcgaga tccattgtgc tggatccgcc accatggagc
     1801 cggaagaggg gacgcccttg tggcggctgc agaagctgcc ggccgagctg ggcccgcagc
     1861 ttcttcacaa aataattgat ggcatttgtg gtcgagctta tcctgtgtac caagattatc
     1921 acactgtttg ggaatcagaa gaatggatgc acgttttaga agatattgcc aaatttttca
     1981 aagccatagt tggtaaaaac ttacctgatg aagagatatt tcagcagttg aatcagttga
     2041 attcacttca tcaagaaact atcatgaaat gcgtgaaaag taggaaagat gaaatcaaac
     2101 aggctctgtc aagagaaata gttgctattt cctctgcaca gctacaggat tttgattggc
     2161 aggtaaagct tgcactttcc agtgacaaga ttgctgcatt acgaatgcca cttttaagcc
     2221 tgcatctaga tgtaaaagaa aatggtgaag taaaacctta ttctattgaa atgagtagag
     2281 aggagctgca gaatctaata cagtccttgg aagcagcgaa taaggtggtc ctgcagttga
     2341 aagcggccgc cgactacaaa gacgatgacg acaagtcact ctgacggccg cgactctaga
     2401 gtgagggtcc ccacctggga cccttgagag tatcaggtct cccacgtggg agacaagaaa
     2461 tccctgttta atatttaaac agcagtgttc cccatctggg tccttgcacc cctcactctg
     2521 gcctcagccg actgcacagc ggcccctgca tccccttggc tgtgaggccc ctggacaagc
     2581 agaggtggcc agagctggga ggcatggccc tggggtccca cgaatttgct ggggaatctc
     2641 gtttttcttc ttaagacttt tgggacatgg tttgactccc gaacatcacc gacgtgtctc
     2701 ctgtttttct gggtggcctc gggacacctg ccctgccccc acgagggtca ggactgtgac
     2761 tctttttagg gccaggcagg tgcctggaca tttgccttgc tggacgggga ctggggatgt
     2821 gggagggagc agacaggagg aatcatgtca ggcctgtgtg tgaaaggaag ctccactgtc
     2881 accctccacc tcttcacccc ccactcacca gtgtcccctc cactgtcaca ttgtaactga
     2941 acttcaggat aataaagtgt ttgcctccaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3001 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaga
     3061 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     3121 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3181 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     3241 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3481 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3541 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3601 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     3661 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3721 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3781 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3841 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3901 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3961 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4441 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4501 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4741 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4801 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     4861 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     4921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     4981 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5161 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5221 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5281 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5341 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5401 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5461 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5521 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5581 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5641 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5701 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5761 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5821 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     5881 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag c
//
