LOCUS       sequence-380105-        4902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian transient expression - expression of Lifeact-Halo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4902)
  AUTHORS   Herron JC, Hu S, Watanabe T, Nogueira AT, Liu B, Kern ME, Aaron J, 
            Taylor A, Pablo M, Chew TL, Elston TC, Hahn KM
  TITLE     Actin nano-architecture of phagocytic podosomes.
  JOURNAL   Nat Commun. 2022 Jul 27;13(1):4363. doi: 10.1038/s41467-022-32038-0.
  PUBMED    35896550
REFERENCE   2  (bases 1 to 4902)
  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 
            Commun."; date: "2022-07-27"; volume: "13(1):4363. doi"; pages: " 
            10.1038/s41467-022-32038-0"
COMMENT     Sequence Label: Lifeact-Halo
FEATURES             Location/Qualifiers
     source          1..4902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..653
                     /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"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             675..1565
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     polyA_signal    1690..1811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1727..1746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1781..1800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1818..2273)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1955..1976)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2167..2186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2300..2404
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2406..2763
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2406..2597
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2429..2449)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2614..2749
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2798..3592
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2798..3592
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2852..2871)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3462..3481
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3780..3799)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3824..3871
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4200..4788
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4689..4708
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatgggcgt ggccgacttg atcaagaagt tcgagtccat ctccaaggag gagggggatc
      661 caccggtcgc caccatggca gaaatcggta ctggctttcc attcgacccc cattatgtgg
      721 aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc
      781 tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg
      841 caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag
      901 acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg
      961 gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg
     1021 ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc
     1081 cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg
     1141 acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg
     1201 gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc
     1261 ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag
     1321 cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga
     1381 agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg
     1441 ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag
     1501 aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtcgacg ctcgagattt
     1561 ccggctaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     1621 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1681 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1741 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1801 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1861 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1921 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1981 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2041 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2101 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2161 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2221 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2281 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2341 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2401 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2461 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2521 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2581 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2641 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2701 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2761 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2821 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2881 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2941 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3001 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3061 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3121 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3181 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3241 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3301 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3361 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3421 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3481 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3541 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3601 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3661 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3721 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     3781 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3841 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3901 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3961 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4021 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4081 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4141 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4201 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4261 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4321 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4381 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4441 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4501 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4561 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4621 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4681 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4741 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4801 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4861 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
