LOCUS       sequence-376732-        5803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5803)
  AUTHORS   Li J, Brieher WM, Scimone ML, Kang SJ, Zhu H, Yin H, von Andrian UH,
            Mitchison T, Yuan J
  TITLE     Caspase-11 regulates cell migration by promoting 
            Aip1-Cofilin-mediated actin depolymerization.
  JOURNAL   Nat Cell Biol. 2007 Mar . 9(3):276-86.
  PUBMED    17293856
REFERENCE   2  (bases 1 to 5803)
  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: "Nat Cell 
            Biol. 2007 Mar . 9(3):276-86."
COMMENT     Sequence Label: PCMV-flag-Caspase11
FEATURES             Location/Qualifiers
     source          1..5803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     298..319
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     582..604
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     596..613
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     597..613
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        774..1153
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1154..1357
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1308..1328
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1387..1452
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      1718..1727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2600..3222
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2799..2818)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3246..3266)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3251..3580
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3431..3566
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3618..3637)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3619..3637)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3651..3667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3651..3667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3664..3686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3675..3691
                     /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)."
     promoter        complement(join(3699..3705,3706..3723,3724..3729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3744..3765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3882..3899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4053..4641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4133..4152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4812..5603,5604..5672))
                     /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     5435..5454
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5673..5777)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
      301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
      361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
      421 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
      481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
      541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
      601 aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt
      661 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
      721 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
      781 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      841 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      901 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      961 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     1021 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     1081 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     1141 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     1201 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     1261 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     1321 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaatta
     1381 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     1441 gacgatgaca agcttgcggc cgcgaattca atggctgaaa acaaacaccc tgacaaacca
     1501 cttaaggtgt tggaacagct gggcaaagaa gtccttacgg agtacctaga aaaattagta
     1561 caaagcaatg tactgaaatt aaaggaggaa gataaacaaa aatttaacaa tgctgaacgc
     1621 agtgacaagc gttgggtttt tgtagatgcc atgaaaaaga aacacagcaa agtaggtgaa
     1681 atgcttctcc agacattctt cagtgtggac ccaggcagcc accatggtga agctaatctg
     1741 gaaatggagg aaccagaaga atcattgaac actctcaagc tttgttcccc tgaagagttc
     1801 acaaggcttt gcagagaaaa gacacaagaa atttacccaa taaaggaggc caatggccgt
     1861 acacgaaagg ctcttatcat atgcaataca gagttcaaac atctctcact gaggtatggg
     1921 gctaaatttg acatcattgg tatgaaaggc cttcttgaag acttaggcta cgatgtggtg
     1981 gtgaaagagg agcttacagc agagggcatg gagtcagaga tgaaagactt tgctgcactc
     2041 tcagaacacc agacatcaga cagcacattc ctggtgctaa tgtctcatgg cacactgcat
     2101 ggcatttgtg gaacaatgca cagtgaaaaa actccagatg tgctacagta tgataccatc
     2161 tatcagatat tcaacaattg ccactgtcca ggtctacgag acaaacccaa agtcatcatt
     2221 gtgcaggcct gcagaggtgg gaactctgga gaaatgtgga tcagagagtc ttcaaaaccc
     2281 cagttgtgca gaggtgtaga tctacctagg aatatggaag ctgatgctgt caagctgagc
     2341 cacgtggaga aggacttcat tgccttctac tctacaaccc cacatcactt gtcctaccga
     2401 gacaaaacag gaggctctta cttcatcact agactcattt cctgcttccg gaaacatgct
     2461 tgctcttgtc atctctttga tatattcctg aaggtgcaac aatcatttga aaaggcaagt
     2521 attcattccc agatgcccac cattgatcgg gcaaccttga caagatattt ctacctcttt
     2581 cctggcaact gaggatcccg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     2641 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     2701 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     2761 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     2821 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     2881 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     2941 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     3001 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     3061 tcccttccct gtccttctga ttttaaaata actataccag caggaggacg tccagacaca
     3121 gcataggcta cctggccatg cccaaccggt gggacatttg agttgcttgc ttggcactgt
     3181 cctctcatgc gttgggtcca ctcagtagat gcctgttgaa ttgggtacgc ggccagcttg
     3241 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3301 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3361 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3421 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3481 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3541 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     3601 aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     3661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3781 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3841 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3901 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3961 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4021 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4081 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4141 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4201 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4261 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4321 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4381 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4441 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4501 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4561 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4621 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4681 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4741 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4801 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4861 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4921 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4981 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5041 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5101 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5161 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5221 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5281 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5341 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5401 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5461 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5521 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5581 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5641 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5701 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5761 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
