LOCUS       sequence-359999-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bi-directional expression vector of monomers with plasma membrane 
            localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  AUTHORS   Petrich A, Dunsing V, Bobone S, Chiantia S
  TITLE     Influenza A M2 recruits M1 to the plasma membrane: A fluorescence 
            fluctuation microscopy study.
  JOURNAL   Biophys J. 2021 Dec 21;120(24):5478-5490. doi: 
            10.1016/j.bpj.2021.11.023. Epub 2021 Nov 19.
  PUBMED    34808098
REFERENCE   2  (bases 1 to 7094)
  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: 
            "10.1016/j.bpj.2021.11.023"; journalName: "Biophys J"; date: 
            "2021-12-21- 21"; volume: "120"; issue: "24"; pages: "5478-5490"
COMMENT     Sequence Label: BiDi-[mp-mCherry2]mp-mEGFP
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    28..252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(278..985)
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     822..840
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      982..991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1758..1767
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1764..1766,1767..1769,1770..2480)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1809..1830)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2070..2089)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2417..2438
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2522..2746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2792..3220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2879..2898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3089..3110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3229..3249)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3234..3563
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3414..3549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3476..3495
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3612..4637
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3612..4637
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4767..4888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4804..4823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4858..4877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4937..4953)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4937..4953)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4950..4972)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4961..4977
                     /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(4985..4991,4992..5009,5010..5015))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5030..5051
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5168..5185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5339..5927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5419..5438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6098..6889,6890..6958))
                     /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     6721..6740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6959..7063)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctgcc tcagaagcca tagagcccac cgcatcccca gcatgcctgc
       61 tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc agaatagaat
      121 gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg acagtgggag
      181 tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat ggctggcaac
      241 tagaaggcac agtcgaggct gatcagcaat tgaattcctt gtacagctcg tccatgccgc
      301 cggtggagtg gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt
      361 gggaaaggat gtccaacttg atgtcgacgt tgtaggcgcc gggcagctgc acgggcttct
      421 tggccttgta ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc
      481 tctgcttgat ctcgcccttc agggcgccgt cctcggggta catccgctca gtggaggcct
      541 cccagcccat ggtacgacac tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca
      601 gcttcacctt gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca
      661 cgccgccgtc ctcgaagttc atcacgcgct cccaattgaa gccctcgggg aaggacagct
      721 tcaagtagtc ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact
      781 gaggggacag gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg
      841 cggtctgggt gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc
      901 cgttcacgga gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt
      961 tatcctcctc gcccttgctc accatggtgg cgaccggttc atcgtcattg agattgtctt
     1021 tcctttttga tttaatacat cccatggatc cgattctagt gaacctccca gcgaagaccg
     1081 cccgttcgac ttctctccca ctgctcctcc ccagtgcccc cgggttgcgc cctcagcggg
     1141 ggcacgataa cgagtggagg caggctacct gagcgagacg cctgggccat ggggcggcgg
     1201 gtccagaacg aacctaacga ttcccacttc cctgcacgga ccaacgcccg ccgcagctcg
     1261 gacttcgccc catcgcaaga gccgttttct ccagtccggg agtcgcgggg accttcatgg
     1321 actctctcgt gctccgtaat gggaggcttc tgcccctaac aaacatggcc gcccactctg
     1381 tcccaccccg ccctttggct ggtatcaaat actagactac ctttcatttc cgggtcactg
     1441 tagatgcgat ggcgccgatt ccaaagactg tggggcggat caagctaggt gagcggtatg
     1501 acaagaccag tgggggtcta ggagggcaga ggcagcccct cttgggtcca cctgtggtct
     1561 gaacgtttcg agtaggattt tatgcagagg gcggagagtc gagggtagag tggggaggac
     1621 tctagcccat ctgtcccgag cccgctgacg aggctccgcc ttcctttcct gtagactgct
     1681 ctctacggcc ctagatcggt accatgggat gtattaaatc aaaaaggaaa gacaatctca
     1741 atgacgatga accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1801 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1861 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1921 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1981 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2041 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2101 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2161 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2221 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2281 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2341 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2401 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2461 gcatggacga gctgtacaag gagctctaga gggcccgttt aaacccgctg atcagcctcg
     2521 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2761 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2821 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2881 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2941 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3001 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3061 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3121 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3181 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3241 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3301 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3421 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3481 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3541 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3601 atcagcacgt gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     3661 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     3721 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     3781 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     3841 tgcttgacat tggggaattt agcgagagcc tgacctattg catctcccgc cgtgcacagg
     3901 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     3961 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     4021 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     4081 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     4141 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     4201 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     4261 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     4321 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     4381 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     4441 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     4501 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     4561 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     4621 cgagggcaaa ggaatagcac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4681 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4741 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4861 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4921 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4981 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5041 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5101 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5761 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5941 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6001 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7081 tgccacctga cgtc
//
