LOCUS       sequence-360008-        6981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bi-directional expression vector of monomers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  AUTHORS   Petazzi RA, Koikkarah AA, Tischler ND, Chiantia S
  TITLE     Detection of Envelope Glycoprotein Assembly from Old-World 
            Hantaviruses in the Golgi Apparatus of Living Cells.
  JOURNAL   J Virol. 2020 Nov 25. pii: JVI.01238-20. doi: 10.1128/JVI.01238-20.
  PUBMED    33239451
REFERENCE   2  (bases 1 to 6981)
  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: "J Virol. 
            2020 Nov 25. pii: JVI.01238-20. doi: 10.1128/JVI.01238-20."
COMMENT     Sequence Label: BiDi-[mEGFP]mCherry2
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(90..110)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        95..424
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      275..410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     337..356
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             473..1498
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             473..1498
                     /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    1628..1749
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1665..1684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1719..1738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1798..1814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1798..1814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1811..1833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1822..1838
                     /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(1846..1852,1853..1870,1871..1876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1891..1912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2029..2046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2200..2788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2280..2299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2959..3750,3751..3819))
                     /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     3582..3601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3820..3924)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3983..4207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(4233..4943)
                     /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     4780..4798
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             join(5606..5608,5609..5611,5612..6322)
                     /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(5651..5672)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5912..5931)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6259..6280
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6364..6588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6634..6981,1..81)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6721..6740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6931..6952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
       61 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
      121 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      181 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      241 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      301 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
      361 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      421 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgatgaaaaa
      481 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
      541 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
      601 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
      661 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
      721 tagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
      781 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
      841 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
      901 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
      961 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     1021 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     1081 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     1141 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     1201 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     1261 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     1321 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     1381 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     1441 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     1501 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     1561 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     1621 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1741 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2641 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3961 atcgggagat ctgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     4021 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     4081 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     4141 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     4201 ggcacagtcg aggctgatca gcaattgaat tctcacttgt acagctcgtc catgccgccg
     4261 gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg
     4321 gaaaggatgt ccaacttgat gtcgacgttg taggcgccgg gcagctgcac gggcttcttg
     4381 gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc
     4441 tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcagt ggaggcctcc
     4501 cagcccatgg tacgacactg cattacgggg ccgtcggagg ggaagttggt gccgcgcagc
     4561 ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac ggtcaccacg
     4621 ccgccgtcct cgaagttcat cacgcgctcc caattgaagc cctcggggaa ggacagcttc
     4681 aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta catgaactga
     4741 ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt cagcttggcg
     4801 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     4861 ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat ggccatgtta
     4921 tcctcctcgc ccttgctcac catggtggga tccgattcta gtgaacctcc cagcgaagac
     4981 cgcccgttcg acttctctcc cactgctcct ccccagtgcc cccgggttgc gccctcagcg
     5041 ggggcacgat aacgagtgga ggcaggctac ctgagcgaga cgcctgggcc atggggcggc
     5101 gggtccagaa cgaacctaac gattcccact tccctgcacg gaccaacgcc cgccgcagct
     5161 cggacttcgc cccatcgcaa gagccgtttt ctccagtccg ggagtcgcgg ggaccttcat
     5221 ggactctctc gtgctccgta atgggaggct tctgccccta acaaacatgg ccgcccactc
     5281 tgtcccaccc cgccctttgg ctggtatcaa atactagact acctttcatt tccgggtcac
     5341 tgtagatgcg atggcgccga ttccaaagac tgtggggcgg atcaagctag gtgagcggta
     5401 tgacaagacc agtgggggtc taggagggca gaggcagccc ctcttgggtc cacctgtggt
     5461 ctgaacgttt cgagtaggat tttatgcaga gggcggagag tcgagggtag agtggggagg
     5521 actctagccc atctgtcccg agcccgctga cgaggctccg ccttcctttc ctgtagactg
     5581 ctctctacgg ccctagatcg gtaccatggt gagcaagggc gaggagctgt tcaccggggt
     5641 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     5701 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     5761 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     5821 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     5881 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     5941 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     6001 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     6061 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
     6121 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     6181 ccccgtgctg ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc
     6241 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     6301 cggcatggac gagctgtaca aggagctcta gagggcccgt ttaaacccgc tgatcagcct
     6361 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6421 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6481 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6541 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     6601 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     6661 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6721 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6781 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6841 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6901 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6961 tcaaccctat ctcggtctat t
//
