LOCUS       sequence-329729-        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  labels F-actin in macrophages.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  AUTHORS   Barros-Becker F, Lam PY, Fisher R, Huttenlocher A
  TITLE     Live imaging reveals distinct modes of neutrophil and macrophage 
            migration within interstitial tissues.
  JOURNAL   J Cell Sci. 2017 Nov 15;130(22):3801-3808. doi: 10.1242/jcs.206128. 
            Epub 2017 Sep 28.
  PUBMED    28972134
REFERENCE   2  (bases 1 to 6094)
  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: 
            "10.1242/jcs.206128"; journalName: "J Cell Sci"; date: "2017-11-15- 
            15"; volume: "130"; issue: "22"; pages: "3801-3808"
COMMENT     Sequence Label: Tg(mpeg1:LifeAct-mRuby)
FEATURES             Location/Qualifiers
     source          1..6094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2093..2114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(2510..2521,2522..2659)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     regulatory      4554..4563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4560..4610
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             complement(4624..4641)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     polyA_signal    5447..5568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5484..5503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5538..5557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(5605..5792,5793..5804)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(5859..5879)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5859..5877)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5898..5914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5898..5914)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5911..5933)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5922..5938
                     /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(5946..5952,5953..5970,5971..5976))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5991..6012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2281 gccggccaca atgacggcta ctacatggtg ccattccttc ctctttatag gaatggagac
     2341 tacctcctgt ccaacaatgc tcttggatac gagtacgcct acctgttgga cccaggtcat
     2401 tgcacaacac cagaaatgcc ctctgatctg caaaagacgt gaatatctgt tcagacaccc
     2461 atatccactc tgttccacac aggtcagagg tttgtccagg agttcttgac agaggtgtaa
     2521 aaagtactca aaaattttac tcaagtgaaa gtacaagtac ttagggaaaa ttttactcaa
     2581 ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa aagtaaaaaa gtactccatt
     2641 aaaattgtac ttgagtatta aggaagtaaa aggctagcaa gatctgctcg agtgttggag
     2701 cacatctgac atctgaaatt atttgaaagc agcagaaatg tttttacagc tattacaaaa
     2761 tttacttaac aagaatcctt taaaatatga caatttattg ccaataattt acagtaagtt
     2821 ttgcagtact ttaatattac aaattataca actgtgctgg cacacaaaaa ataacttaat
     2881 ccagcacaac aacaacaaca acaactacaa aaagcatatt tccttttgct ttttcttgtt
     2941 cactcaactt ttggaaacat caactgcact gacctaaaat tatacagtat gtcagtaata
     3001 caaaaacact tagttaggtt aacataagat tattagtttt ctggagaggt gaactactct
     3061 gtaagaaacc tttttaagtt gtgccaacta aacattttta tcagcagaac tacaatgctt
     3121 gatgagtgaa caaaaaaaca aacaaacctt atttcctttt gttttttctt gtgtacttaa
     3181 catttttttc acctttattt gaaacgacag tagggagtat ggacaggaaa gtgttgggag
     3241 aggagaggga tgagcataga accaggaggt aggaatcaaa ctcaggtcac tatgaacacc
     3301 ggagtgccag gtgtcaacac actaaccact acaccatcaa ctccaacatt tacttgaaaa
     3361 catcagttga gtgaacaaaa aactcttgtg gaaattggta ctgagaaata atacattatc
     3421 acaacaaaca aaacagagac aaatgttaac aaaacaatta ttagaaacag aattagaaac
     3481 cggacaggag gtaaaacaat aatgataaac aagacaagga tcaaaacatg gcaggacaaa
     3541 gacagttttt tgttcactca acctcagata ttttaggtca gtgcaactga tgtttacaaa
     3601 ggttgtatgc atgtcaacac caatggtgta gtgattagtg tgttgacaca aggcactcca
     3661 gtgttcacac tgatgctggt ttgattccca catcgatgtc ctttgctaat cctgcctctc
     3721 tccccatgct ttcctgtcta tactctctac tgttgtgtca attaaaggtg aaaaaagtta
     3781 agcacaaaag aaaaatctaa ggatataagg attatgtttt tttttgttca ctcaaaatca
     3841 ggcattccag ttctgcagac aaaaaatgtt cagttggcac aacttaaaaa gttttgttac
     3901 agagtagttc acctctccag aaaactaata atcttatgtt aacccaacta aatgttttta
     3961 ctgaaaaata attgaatgtc agtgcagctg atgttttttt aaaagtggag agaacaaaaa
     4021 aagtgaaagg aaataaggat tcattgttgt accgacttat ttttttgaag aaaattctgc
     4081 tatagggctt tcatgatccc ctatgctcac agcagtttta ggagatgctt ataataaaga
     4141 ggatttgaaa aacctgctaa tcttgtacat ttctaaaaca agaagccaac caatttcaat
     4201 tcctcctctt tcaaagccaa tggaaaattt aaaacgcaca gtctcttgcg tcatcaaaac
     4261 cattctgaag actcatcaaa atccttcgta gctttaaata taaatctatt attgaacaga
     4321 tcatacttca caaaaaatat aaatatattt gattcattca cttcatgaaa ttgtccttca
     4381 acttaacgtt tgaacgcaag gtaccaaaac acaaagagga acctcatagt ctcatgaccc
     4441 ttcccccacg gatttgaaac tgagaagtgg ttttattgag gaactcagca aaatctataa
     4501 tcaccatatt tgattgaaaa tcacattagt gcaggagaca gcaaaaggat ctcgccacca
     4561 tgggtgtcgc agatttgatc aagaaattcg aaagcatctc aaaggaagaa ggggatccgg
     4621 gtgctggtgc tggtgctggt gctatgggca agcttaccat ggccagctcc gaggatgtca
     4681 tcaaagagtt tatgagattt aaggtcaaga tggagggaag cgtcaacgga cacgagttcg
     4741 agattgaggg agaaggagaa ggccggcctt acgagggcac acaaaccgct aagctcaagg
     4801 tcacaaaagg aggacccctc cccttctcct gggatattct gagccctcag ttccagtacg
     4861 gaagcaaagc ctatgttaaa caccctgccg acatccctga ctatctgaag ctctccttcc
     4921 ctgaaggctt caagtgggag agattcatga acttcgagga cggaggcgtg gtgacagtca
     4981 cacaagatag caccctccag gacggagagt ttatttataa ggtgaaactc agaggaacca
     5041 acttcccctc cgatggccct gtcatgcaaa aaaaaacaat gggatgggaa gcctccaccg
     5101 agagaatgta tcctgaggat ggcgctctga aaggcgaaat taaaatgaga ctgaaactca
     5161 aagacggagg acactacgat gccgaggtca aaacaaccta caaggccaag aaacaagtgc
     5221 agctgcctgg cgcctacatg actgatatta aactcgacat tatcagccat aatggggact
     5281 acaccatcgt ggaacaatat gagagagctg agggcagaca tagcacaggc gcttaatcta
     5341 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5401 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5461 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5521 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgagatcct
     5581 gcggccgcaa gggaaaatag aatgaagtga tctccaaaaa ataagtactt tttgactgta
     5641 aataaaattg taaggagtaa aaagtacttt tttttctaaa aaaatgtaat taagtaaaag
     5701 taaaagtatt gatttttaat tgtactcaag taaagtaaaa atccccaaaa ataatactta
     5761 agtacagtaa tcaagtaaaa ttactcaagt actttacacc tctggttctt gaccccctac
     5821 cttcagcaag cccagcagat ccactagatc ttttgttccc tttagtgagg gttaattgcg
     5881 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5941 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6001 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6061 cagctgcatt aatgaatcgg ccaacgcgcg ggga
//
