LOCUS       sequence-360014-        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-mEGFP]mp-mCherry2
FEATURES             Location/Qualifiers
     source          1..7094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    35..259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(285..995,996..998,999..1001))
                     /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(327..348)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     676..695
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     935..956
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      998..1007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1774..1783
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1780..2487
                     /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     complement(1925..1943)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    2529..2753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2799..3227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2886..2905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3096..3117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3236..3256)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3241..3570
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3421..3556
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3483..3502
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3619..4644
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3619..4644
                     /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    4774..4895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4811..4830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4865..4884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4944..4960)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4944..4960)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4957..4979)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4968..4984
                     /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(4992..4998,4999..5016,5017..5022))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5037..5058
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5175..5192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5346..5934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5426..5445)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6105..6896,6897..6965))
                     /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     6728..6747
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6966..7070)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacgtcgac ggatcgggag atctgcctca gaagccatag agcccaccgc atccccagca
       61 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
      121 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
      181 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
      241 tggcaactag aaggcacagt cgaggctgat cagcaattga attccttgta cagctcgtcc
      301 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
      361 tcgttggggt ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc
      421 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
      481 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
      541 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
      601 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
      661 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
      721 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
      781 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
      841 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
      901 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
      961 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcgac cggttcatcg
     1021 tcattgagat tgtctttcct ttttgattta atacatccca tggatccgat tctagtgaac
     1081 ctcccagcga agaccgcccg ttcgacttct ctcccactgc tcctccccag tgcccccggg
     1141 ttgcgccctc agcgggggca cgataacgag tggaggcagg ctacctgagc gagacgcctg
     1201 ggccatgggg cggcgggtcc agaacgaacc taacgattcc cacttccctg cacggaccaa
     1261 cgcccgccgc agctcggact tcgccccatc gcaagagccg ttttctccag tccgggagtc
     1321 gcggggacct tcatggactc tctcgtgctc cgtaatggga ggcttctgcc cctaacaaac
     1381 atggccgccc actctgtccc accccgccct ttggctggta tcaaatacta gactaccttt
     1441 catttccggg tcactgtaga tgcgatggcg ccgattccaa agactgtggg gcggatcaag
     1501 ctaggtgagc ggtatgacaa gaccagtggg ggtctaggag ggcagaggca gcccctcttg
     1561 ggtccacctg tggtctgaac gtttcgagta ggattttatg cagagggcgg agagtcgagg
     1621 gtagagtggg gaggactcta gcccatctgt cccgagcccg ctgacgaggc tccgccttcc
     1681 tttcctgtag actgctctct acggccctag atcggtacca tgggatgtat taaatcaaaa
     1741 aggaaagaca atctcaatga cgatgaaccg gtcgccacca tggtgagcaa gggcgaggag
     1801 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     1861 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     1921 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     1981 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2041 ttgaagctgt ccttccccga gggcttcaat tgggagcgcg tgatgaactt cgaggacggc
     2101 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2161 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagtgtcg taccatgggc
     2221 tgggaggcct ccactgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2281 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2341 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtcg acatcaagtt ggacatcctt
     2401 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2461 accggcggca tggacgagct gtacaaggag ctctagaggg cccgtttaaa cccgctgatc
     2521 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2581 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2641 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2701 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2761 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2821 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2881 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2941 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3001 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3061 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3121 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3181 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3421 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3481 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3541 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3601 ggatctgatc agcacgtgat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt
     3661 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     3721 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     3781 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
     3841 ccggaagtgc ttgacattgg ggaatttagc gagagcctga cctattgcat ctcccgccgt
     3901 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     3961 gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc
     4021 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     4081 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     4141 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     4201 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     4261 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     4321 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     4381 gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     4441 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     4501 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg
     4561 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     4621 actcgtccga gggcaaagga atagcacgtg ctacgagatt tcgattccac cgccgccttc
     4681 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4741 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4801 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4861 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4921 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4981 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5041 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5101 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5161 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5221 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5281 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5341 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5401 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5461 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5521 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5581 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5641 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5701 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5761 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5821 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5881 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5941 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6001 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6061 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6121 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6181 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6241 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6301 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6361 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6421 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6481 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6541 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6601 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6661 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6721 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6781 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6841 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6901 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6961 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7021 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7081 cgaaaagtgc cacc
//
