LOCUS       sequence-194461-        8345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses COPZ1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8345)
  AUTHORS   Galligan JT, Martinez-Noel G, Arndt V, Hayes S, Chittenden TW, 
            Harper JW, Howley PM
  TITLE     Proteomic analysis and identification of cellular interactors of the
            giant ubiquitin ligase HERC2.
  JOURNAL   J Proteome Res. 2015 Feb 6;14(2):953-66. doi: 10.1021/pr501005v. 
            Epub 2014 Dec 15.
  PUBMED    25476789
REFERENCE   2  (bases 1 to 8345)
  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: "10.1021/pr501005v";
            journalName: "J Proteome Res"; date: "2015-02-6- 6"; volume: "14"; 
            issue: "2"; pages: "953-66"
COMMENT     Sequence Label: p7055 pHAGE-P-CMVt-N-HA-GAW-COPZ1
FEATURES             Location/Qualifiers
     source          1..8345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..611
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     512..531
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     765..782
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1022..1043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1058..1063,1064..1081,1082..1088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1096..1112
                     /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     1101..1123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1120..1136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1120..1136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1186..1205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1397..2030
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2077..2202
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2699..2932
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3117..3161
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    3454..3571
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3623..3926
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3927..4130
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4081..4101
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4132..4150
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4153..4171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4181..4205
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4292..4301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4304..4330
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4346..4370
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4902..4926)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4945..5444
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4980..5003)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5377..5396
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5465..6064
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5465..5484)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5961..5981
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6131..6719
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6184..6204)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6602..6613)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6794..7027
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7146..7164)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7232..7336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7337..7405,7406..8197)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7555..7574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
       61 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
      181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
      241 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
      301 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
      361 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
      421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
      481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
      541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      661 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      781 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      841 aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
      901 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
      961 aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     1021 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     1081 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgacatga
     1141 ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1201 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
     1261 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
     1321 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
     1381 ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga tatccttgat
     1441 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     1501 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     1561 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     1621 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     1681 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     1741 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     1801 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     1861 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     1921 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     1981 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     2041 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     2101 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     2161 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     2221 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     2281 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     2341 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     2401 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     2461 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     2521 aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga ggagatatga
     2581 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     2641 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     2701 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     2761 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     2821 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     2881 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     2941 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     3001 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     3061 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     3121 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     3181 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     3241 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     3301 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     3361 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc
     3421 gccgaattca caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     3481 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     3541 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     3601 agtttggact agtggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     3661 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     3721 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     3781 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     3841 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     3901 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     3961 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4021 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4081 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
     4141 gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc
     4201 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     4261 ccagcctccg gactctagcg tttaaacgct agccaccatg gcgtacccat acgatgttcc
     4321 agattacgct gctcgaggtt atcaaacaag tttgtacaaa aaagttggca tggaggcgct
     4381 gattttggaa ccttccctgt atactgtcaa agccatcctg attctggaca atgatggaga
     4441 tcgacttttt gccaagtact atgacgacac ctaccccagt gtcaaggagc aaaaggcctt
     4501 tgagaagaac attttcaaca agacccatcg gactgacagt gaaattgccc tcttggaagg
     4561 cctgacagtg gtatacaaaa gcagtataga tctctatttc tatgtgattg gcagctccta
     4621 tgaaaatgag ctgatgctta tggctgttct gaactgtctc ttcgactcat tgagccagat
     4681 gctgaggaaa aatgtagaaa agcgagcact gctggagaac atggaggggc tgttcttggc
     4741 tgtggatgaa attgtagatg gaggggtgat cctagagagt gatccccagc aggtggtaca
     4801 ccgggtggca ttaaggggtg aagatgtccc ccttacggag cagaccgtgt ctcaggtgct
     4861 gcagtcagcc aaagaacaga tcaagtggtc actccttcgg tgcccaactt tcttgtacaa
     4921 agtggttcga taacgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     4981 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     5041 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     5101 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     5161 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     5221 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     5281 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     5341 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     5401 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct
     5461 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     5521 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     5581 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5641 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5701 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5761 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5821 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5881 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5941 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     6001 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     6061 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     6121 gatagatcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6181 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6241 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6301 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6361 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6421 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     6481 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     6541 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     6601 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     6661 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     6721 tgagatcctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6781 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     6841 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     6901 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     6961 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7021 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7081 atatcagaga gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg
     7141 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     7201 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7261 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     7321 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     7381 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     7441 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     7501 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     7561 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7621 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7681 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7741 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7801 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7861 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7921 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7981 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     8041 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     8101 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     8161 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     8221 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     8281 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     8341 ccccg
//
