LOCUS       sequence-353434-        4607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of ZZ-TEV-MZT2-3C-mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4607)
  AUTHORS   Wieczorek M, Ti SC, Urnavicius L, Molloy KR, Aher A, Chait BT, 
            Kapoor TM
  TITLE     Biochemical reconstitutions reveal principles of human gamma-TuRC 
            assembly and function.
  JOURNAL   J Cell Biol. 2021 Mar 1;220(3). pii: 211719. doi: 
            10.1083/jcb.202009146.
  PUBMED    33496729
REFERENCE   2  (bases 1 to 4607)
  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 Cell 
            Biol."; date: "2021-03-1"; volume: "220(3). pii: 211719. doi"; 
            pages: " 10.1083/jcb.202009146"
COMMENT     Sequence Label: pACEBac1-ZZ-TEV-MZT2-3C-mEGFP
FEATURES             Location/Qualifiers
     source          1..4607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      190..778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     679..698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  913..1136
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1203..1736)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1925..1930,1931..1947,1948..1953))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(2024..2057)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2134..2225
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2150..2167
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2357..2530
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAYYEILHLPNLNEGQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             2531..2704
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAEKLNDAQAPK"
     CDS             2762..2782
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2804..2824
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3314..3337
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             join(3338..3340,3341..3343,3344..4057)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3383..3404)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3644..3663)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3991..4012
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4187..4321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4224..4243)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4278..4297
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4442..4607
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 accggttgac ttgggtcaac tgtcagacca agtttactca tatatacttt agattgattt
       61 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
      121 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
      181 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
      241 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      301 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      361 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      421 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      481 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      541 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      601 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      661 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      721 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      781 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      841 ctttcctgcg ttatcccctg attgacttgg gtcgctcttc ctgtggatgc gcagatgccc
      901 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
      961 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     1021 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     1081 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tgtaaagact atattcgcgg
     1141 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     1201 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     1261 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     1321 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     1381 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     1441 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     1501 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     1561 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     1621 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     1681 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt
     1741 gctgctccat aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg
     1801 aggcatagac tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     1861 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     1921 ttacagttta cgaaccgaac aggcttatgt caactgggtt cgtgccttca tccgtttcca
     1981 cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc gccataactt cgtatagcat
     2041 acattatacg aagttatctg taactataac ggtcctaagg tagcgagttt aaacactagt
     2101 atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa aatgataacc
     2161 atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta
     2221 taaatattcc ggattattca taccgtccca ccatcgggcg cggatcccgg tccgaagcgc
     2281 gcggaattca aaggcctacg tcgacgagct cacttgtcgc ggccgcatgg caggccttgc
     2341 gcaacacgat gaagccgtgg acaacaaatt caacaaagaa caacaaaacg cgtactatga
     2401 gatcttacat ttacctaact taaacgaagg acaacgaaac gccttcatcc aaagtttaaa
     2461 agatgaccca agccaaagcg ctaacctttt agcagaagct aaaaagctaa atgatgctca
     2521 ggcgccgaaa gtagacaaca aattcaacaa agaacaacaa aacgcgttct atgagatctt
     2581 acatttacct aacttaaacg aagaacaacg aaacgccttc atccaaagtt taaaagatga
     2641 cccaagccaa agcgctaacc ttttagcaga agctgaaaag ctaaatgatg ctcaggcgcc
     2701 gaaagtagac gcaaattccg cggggaagtc aaccgattat gatattccaa ctactgctag
     2761 cgagaaccta tatttccagg gagattatga tatccctacg actgagaatc tttattttca
     2821 gggcgaattc atggcggcgc agggcgtagg gcctgggccg gggtcggcgg cgcccccggg
     2881 gctggaggcg gcccggcaga agctggcgct gcggcgcaag aaggtgctga gcaccgagga
     2941 gatggagctg tacgagctgg ctcaggcggc gggcggcggt atcgaccccg acgtgttcaa
     3001 gatcctggtg gacctgctga agctgaacgt ggcccccctc gccgtcttcc agatgctcaa
     3061 gtccatgtgt gccgggcaga ggctagcgag cgagccccag gaccctgcgg ccgtgtctct
     3121 gcccacgtcg agcgtgcccg agacccgagg gagagacaaa ggcagcgctg ccctcggggg
     3181 agtattggcc ctggcggaac gcagcaacca cgagggatcc agccagagga tgccacgcca
     3241 gcccagcgct accaggctgc ccaagggggg cgggcctggg aagagcccta cgcagggcag
     3301 caccggtctc gagctggagg tgctgttcca gggacctatg gtgagcaagg gcgaggagct
     3361 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     3421 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     3481 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     3541 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     3601 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     3661 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     3721 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     3781 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     3841 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3901 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct
     3961 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4021 cgggatcact ctcggcatgg acgagctgta caagtgaaag cttgtcgaga agtactagag
     4081 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4141 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4201 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4261 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     4321 ctgatcactg cttgagccta gaagatccgg ctgctaacaa agcccgaaag gaagctgagt
     4381 tggctgctgc caccgctgag caataactat cataacccct agggtatacc catctaattg
     4441 gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt tcgtattagc
     4501 ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata atccttaaaa
     4561 actccatttc cacccctccc agttcccaac tattttgtcc gcccaca
//
