pcDNA-LifeAct-mBiyu

LOCUS       pcDNA-LifeAct-mBiyu               6219 bp DNA     circular SYN 17-MAY-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6219)
  AUTHORS   sunwenjuan
  TITLE     Direct Submission
  JOURNAL   Exported Sep 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             901..951
                     /codon_start=1
                     /label=Lifeact
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     complement(921..959)
                     /label=lifeact-backbone-R
     CDS             976..1716
                     /codon_start=1
                     /label=mBiyu
                     /translation="SKGEEDNMASYGALLFREKIPYVVEMEGDVEGMKFSVRGKGHGDA
                     NTGKIEASFICTTGELPVPWSSILTTVTYGAQCFAKYPNDIKDYLKSTMPEGYVQERTI
                     TFENDGIYKTRAEVTYEKGSVYNRVTLNGSGFKKGGNILGKKLEFNFNASRTYVLPDVE
                     NNGIKCYINIRHNVIGGGQIRAEHQQLNTPLGGGPVDIPHYHAIQTHTILSKDPKETRD
                     HMNVVEVFRAIDCKTAYAGMDELYK"
     polyA_signal    1819..2043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2089..2517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2531..2860
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2711..2846
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2927..3721
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3816..3839)
                     /label=lifeact-YU-R
     primer_bind     3822..3852
                     /label=lifeact-backbone-F
     polyA_signal    3895..4016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4065..4081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4089..4105
                     /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(4113..4143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4158..4179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4467..5052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5223..6083)
                     /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"
     promoter        complement(6084..6188)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atgggagtgg ctgacctgat caagaagttc gagagcatca gcaaggagga gggcgatccc
      961 ccagtggctc tggcttccaa aggcgaagag gataatatgg ccagctacgg cgccctgctg
     1021 tttagggaaa agatccccta cgtggtggag atggagggcg acgttgaggg aatgaagttc
     1081 agcgtgaggg gcaagggcca cggagatgct aataccggca agatcgaggc cagcttcatc
     1141 tgtacaacag gcgagctgcc cgtgccttgg tcttctatcc tgacaacagt gacctacggc
     1201 gcccagtgtt ttgccaagta ccccaacgat atcaaggact acctgaagag caccatgcct
     1261 gagggctacg tgcaggagag gaccatcacc ttcgagaacg atggcatcta caagacaaga
     1321 gccgaggtga catacgagaa gggctccgtg tacaacaggg tgacactgaa cggcagcggc
     1381 ttcaagaagg gcggaaacat cctgggcaag aagctggagt ttaatttcaa cgccagcagg
     1441 acctacgtgc tgcctgatgt ggagaataac ggcatcaagt gttacatcaa catcagacac
     1501 aacgtgatcg gcggcggcca gatcagagct gaacaccaac agctgaacac acccctgggc
     1561 ggaggacctg ttgatatccc tcactaccac gccatccaga cccacaccat cctgtccaag
     1621 gaccccaagg agaccagaga tcacatgaat gtggtggagg tgttcagagc catcgattgc
     1681 aagaccgcct acgccggaat ggacgagctg tacaagtgag gatccactag tccagtgtgg
     1741 tggaattctg cagatatcca gcacagtggc ggccgctcga gtctagaggg cccgtttaaa
     1801 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     1861 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     1921 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     1981 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2041 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2101 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2161 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2221 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2281 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2341 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2401 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2461 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2521 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     2581 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     2641 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     2701 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     2761 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     2821 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     2881 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2941 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3001 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3061 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3121 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3181 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3241 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3301 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3361 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3421 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3481 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3541 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3601 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3661 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3721 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3781 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3841 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     3901 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3961 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4021 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4081 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4141 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4261 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4981 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5041 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5161 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5221 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5281 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5341 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5401 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5461 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5521 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5581 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5641 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5701 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5761 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5821 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5881 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5941 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6001 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6061 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6121 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6181 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//