LOCUS       sequence-218697-        5808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of I3 in insect cells (Baculovirus system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5808)
  AUTHORS   Weith M, Seiler J, van den Boom J, Kracht M, Hulsmann J, Primorac I,
            Del Pino Garcia J, Kaschani F, Kaiser M, Musacchio A, Bollen M, 
            Meyer H
  TITLE     Ubiquitin-Independent Disassembly by a p97 AAA-ATPase Complex Drives
            PP1 Holoenzyme Formation.
  JOURNAL   Mol Cell. 2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020.
  PUBMED    30344098
REFERENCE   2  (bases 1 to 5808)
  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: "Mol Cell. 
            2018 Oct 11. pii: S1097-2765(18)30791-3. doi: 
            10.1016/j.molcel.2018.09.020."
COMMENT     Sequence Label: pFL I3
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..128
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     53..70
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    745..879
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(782..801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     836..855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  989..1154
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1338..1792
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1425..1444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1634..1655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1818..1922
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1923..1991,1992..2783)
                     /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(2141..2160)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2954..3542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3443..3462
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3696..3713
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3847..4071
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4138..4671)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4860..4865,4866..4882,4883..4888))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(5284..5317)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5507..5555
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5580..5599
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     promoter        complement(5691..5800)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
ORIGIN
        1 agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat taaaatgata
       61 accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta ataaaaaaac
      121 ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatct acaaccatgg
      181 ccgaggcagg ggctgggctg agcgagaccg tcactgagac aacggttacc gtgacaaccg
      241 agcccgagaa ccggagcctt accatcaaac ttcggaaacg gaagccagag aaaaaggtag
      301 aatggacaag tgacactgtg gacaatgaac acatgggccg ccgctcatcc aaatgctgct
      361 gtatttatga gaaacctcgg gcctttggcg agagctccac ggaaagtgat gaggaggaag
      421 aagagggctg tggtcataca cactgtgtac gtggccaccg caaaggacgg cgtcgtgcaa
      481 ccctaggacc gacccccacc acccctcccc agcctcctga cccttcccag ccccctccag
      541 ggccaatgca gcactaatga gtcgacgagc tcactagtcg cggccgcttt cgaatctaga
      601 gcctgcagtc tcgacaagct tgtcgagaag tactagagga tcataatcag ccataccaca
      661 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
      721 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
      781 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      841 ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct tgagcctaga
      901 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
      961 ataactatca taacccctag gagatccgaa ccagataagt gaaatctagt tccaaactat
     1021 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     1081 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     1141 tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa acatcctgcc
     1201 aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga atgaaaattt
     1261 ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc ttatttgcag
     1321 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1621 cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     1681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     1741 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     1801 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1861 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1921 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1981 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2041 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2101 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2161 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2221 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2281 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2341 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2401 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2461 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2521 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2581 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2641 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2701 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2761 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2821 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2881 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2941 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3001 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3421 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3481 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3541 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3601 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3661 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3721 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     3781 accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat gccctgcgta
     3841 agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa actatgacaa
     3901 taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta tgctgtgaaa
     3961 aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa gtgcaaattg
     4021 cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt cgcggcgttg
     4081 tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta
     4141 ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc ccaactttgt
     4201 atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca cataagcacc
     4261 aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg
     4321 gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct gctcaaactt
     4381 gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc
     4441 gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct gatgttggga
     4501 gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc gcatggattt
     4561 gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc cacctaactt
     4621 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc
     4681 tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca
     4741 tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac
     4801 cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca
     4861 gtttacgaac cgaacaggct tatgtcaact gggtttgtgc cttcatccgt ttccacggtg
     4921 tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga
     4981 acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct
     5041 acggcaaggt gctgtgcacg gatctgccct tgcttcagga gatcggtaga cctcggccgt
     5101 cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg
     5161 aaggcgagca tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacagc
     5221 tttgtttgta ctatcaacag gttgaactgc tgatcaacag atcctctacg cggccgcggt
     5281 accataactt cgtatagcat acattatacg aagttatctg gtttaaacgt acccgtagtg
     5341 gctatggcag ggcttgccgc cccgacgttg gctgcgagcc ctgggccttc acccgaactt
     5401 gggggttggg gtggggaaaa ggaagaaacg cgggcgtatt ggtcccaatg gggtctcggt
     5461 ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac aaacgaccca
     5521 acacccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     5581 tgtctccttc cgtgtttcag ttagcctccc ccatctcccg gtaccgcatg ctatgcatca
     5641 gctgctagca ccatggctcg agatcccggg tgatcaagtc ttcgtcgagt gattgtaaat
     5701 aaaatgtaat ttacagtata gtattttaat taatatacaa atgatttgat aataattctt
     5761 atttaactat aatatattgt gttgggttga attaaaggtc cgtatact
//
