LOCUS       sequence-346330-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  F-actin marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4948)
  TITLE     Gadella lab plasmids
REFERENCE   2  (bases 1 to 4948)
  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"
COMMENT     Sequence Label: LifeAct-HaloTag
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     193..212
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(511..530)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    555..602
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      931..1519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1420..1439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1694..1997
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1998..2201
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2152..2172
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2272..2322
                     /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      2338..2347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3369..3490
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3406..3425)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3460..3479
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3497..3952)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3634..3655)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3846..3865
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3979..4083
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4085..4442
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4085..4276
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4108..4128)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4293..4428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4477..4948,1..323)
                     /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             join(4477..4948,1..323)
                     /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(4531..4550)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
       61 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
      121 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
      181 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
      241 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
      301 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
      361 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
      421 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
      481 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
      541 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
      601 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
      661 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
      721 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
      781 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
      841 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
      901 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      961 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1021 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1081 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1141 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1201 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1261 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1321 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1381 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1441 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1501 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1561 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1621 taccgccatg cattagttat taatagtaat caattacggg gtcattagtt catagcccat
     1681 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     1741 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     1801 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     1861 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     1921 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     1981 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2041 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2101 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2161 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2221 tccgctagcg ctaccggact cagatctcga gctcaagctt cgaattccac catgggcgtg
     2281 gccgacctga tcaagaagtt cgagagcatc agcaaggagg agggggatcc accggtcgcc
     2341 accatggccg agatcggcac cggcttcccc ttcgaccccc actacgtgga ggtgctgggc
     2401 gagcgcatgc actacgttga cgtgggcccc cgcgacggca cccccgtgct gttcctgcac
     2461 ggcaacccca ccagcagcta cgtgtggcgc aacatcatcc cccacgtggc ccccacccac
     2521 cgcactatcg cccccgacct gatcggcatg ggcaagagcg acaagcccga cctgggctac
     2581 ttcttcgacg accacgtgcg cttcatggac gccttcatcg aggccctggg cctggaggag
     2641 gtggtgctgg tgatccacga ctggggcagc gccctgggct tccactgggc caagcgcaac
     2701 cccgagcgcg tgaagggcat cgccttcatg gagttcatcc gccccatccc cacctgggac
     2761 gagtggcccg agttcgcccg cgagaccttc caggccttcc gcaccaccga cgtgggccgc
     2821 aagctgatca tcgaccagaa cgtgttcatc gagggcaccc tgcccatggg cgtggtgcgc
     2881 cccctgaccg aggtggagat ggaccactac cgcgagccct tcctgaaccc cgtggaccgc
     2941 gagcccctgt ggcgcttccc caacgagctg cccatcgccg gcgagcccgc caacatcgtg
     3001 gccctggtgg aggagtacat ggactggctg caccagagcc ccgtgcccaa gctgctgttc
     3061 tggggcaccc ccggcgtgct gatccccccc gccgaggccg cccgcctggc caagagcctg
     3121 cccaacgtta aggccgtgga catcggcccc ggcctgaacc tgctccagga ggacaacccc
     3181 gacctgatcg gcagcgagat cgcccgctgg ctgagcaccc tggagatcag cggcgtgtac
     3241 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3301 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3361 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3421 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3481 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3541 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3601 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3661 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3721 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3781 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3841 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3901 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4081 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4141 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4201 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4261 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4321 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4381 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4441 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4501 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4561 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4621 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4681 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4741 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4801 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4861 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4921 gaagccggtc ttgtcgatca ggatgatc
//
