LOCUS       sequence-353416-        4234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of gamma-tubulin(N229A)-TEV-His6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4234)
  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 4234)
  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-gamma-tubulin(N229A)-TEV-His6
FEATURES             Location/Qualifiers
     source          1..4234
                     /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             3643..3663
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3664..3681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3814..3948
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3851..3870)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3905..3924
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4069..4234
                     /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 tgccgaggga aatcatcacc ctacagttgg gccagtgcgg caatcagatt
     2341 gggttcgagt tctggaaaca gctgtgcgcc gagcatggta tcagccccga gggcatcgtg
     2401 gaggagttcg ccaccgaggg cactgaccgc aaggacgtct ttttctacca ggcagacgat
     2461 gagcactaca tcccccgggc cgtgctgctg gacttggaac cccgggtgat ccactccatc
     2521 ctcaactccc cctatgccaa gctctacaac ccagagaaca tctacctgtc ggaacatgga
     2581 ggaggagctg gcaacaactg ggccagcgga ttctcccagg gagaaaagat ccatgaggac
     2641 atttttgaca tcatagaccg ggaggcagat ggtagtgaca gtctagaggg ctttgtgctg
     2701 tgtcactcca ttgctggggg gacaggctct ggactgggtt cctacctctt agaacggctg
     2761 aatgacaggt atcctaagaa gctggtgcag acatactcag tgtttcccaa ccaggacgag
     2821 atgagcgatg tggtggtcca gccttacaat tcactcctca cactcaagag gctgacgcag
     2881 aatgcagact gtgtggtggt gctggacaac acagccctga accggattgc cacagaccgc
     2941 ctgcacatcc agaacccatc cttctcccag atcgcccagc tggtgtctac catcatgtca
     3001 gccagcacca ccaccctgcg ctaccctggc tacatgaaca atgacctcat cggcctcatc
     3061 gcctcgctca ttcccacccc acggctccac ttcctcatga ccggctacac ccctctcact
     3121 acggaccagt cagtggccag cgtgaggaag actacggtcc tggatgtcat gaggcggctg
     3181 ctgcagccca agaacgtgat ggtgtccaca ggccgagacc gccagaccaa ccactgctac
     3241 atcgccatcc tcaacatcat ccagggagag gtggacccca cccaggtcca caagagcttg
     3301 cagaggatcc gggaacgcaa gttggccaac ttcatcccgt ggggccccgc cagcatccag
     3361 gtggccctgt cgaggaagtc tccctacctg ccctcggccc accgggtcag cgggctcatg
     3421 atggccaacc acaccagcat ctcctcgctc ttcgagagaa cctgtcgcca gtatgacaag
     3481 ctgcgtaagc gggaggcctt cctggagcag ttccgcaagg aggacatgtt caaggacaac
     3541 tttgatgaga tggacacatc cagggagatt gtgcagcagc tcatcgatga gtaccatgcg
     3601 gccacacggc cagactacat ctcctggggc acccaggagc aggaaaacct gtattttcag
     3661 ggccatcacc atcatcacca ctagaagctt gtcgagaagt actagaggat cataatcagc
     3721 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3781 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3841 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3901 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
     3961 gagcctagaa gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     4021 cgctgagcaa taactatcat aacccctagg gtatacccat ctaattggaa ccagataagt
     4081 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     4141 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     4201 ccctcccagt tcccaactat tttgtccgcc caca
//
