LOCUS       sequence-360029-        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-[mCherry2]mEGFP
FEATURES             Location/Qualifiers
     source          1..6981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    321..442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(358..377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     412..431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(491..507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(491..507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(504..526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    515..531
                     /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(539..545,546..563,564..569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    584..605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(722..739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(893..1481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(973..992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1652..2443,2444..2512))
                     /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     2275..2294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2513..2617)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2676..2900
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(2926..3639,3640..3642,3643..3645))
                     /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(2971..2992)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3320..3339
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3579..3600
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4308..5015
                     /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(4453..4471)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    5057..5281
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5327..5755
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5414..5433)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5624..5645
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5764..5784)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5769..6098
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5949..6084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6011..6030
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(6147..6981,1..191)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             join(6147..6981,1..191)
                     /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"
ORIGIN
        1 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
       61 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
      121 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
      181 caaaggaata gcacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
      241 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
      301 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
      361 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      421 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
      481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      541 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      841 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      901 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      961 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1021 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1081 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1141 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1201 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1261 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1321 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1381 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1441 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1501 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1561 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1621 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1681 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     1741 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     1801 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     1861 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     1921 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     1981 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2041 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2101 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2161 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2221 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2281 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2341 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2401 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2461 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2521 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2581 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2641 ctgacgtcga cggatcggga gatctgcctc agaagccata gagcccaccg catccccagc
     2701 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
     2761 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
     2821 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
     2881 ctggcaacta gaaggcacag tcgaggctga tcagcaattg aattctcact tgtacagctc
     2941 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
     3001 cttctcgttg gggtctttgc tcagcttgga ctgggtgctc aggtagtggt tgtcgggcag
     3061 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
     3121 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
     3181 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
     3241 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     3301 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     3361 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     3421 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     3481 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     3541 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     3601 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg gatccgattc
     3661 tagtgaacct cccagcgaag accgcccgtt cgacttctct cccactgctc ctccccagtg
     3721 cccccgggtt gcgccctcag cgggggcacg ataacgagtg gaggcaggct acctgagcga
     3781 gacgcctggg ccatggggcg gcgggtccag aacgaaccta acgattccca cttccctgca
     3841 cggaccaacg cccgccgcag ctcggacttc gccccatcgc aagagccgtt ttctccagtc
     3901 cgggagtcgc ggggaccttc atggactctc tcgtgctccg taatgggagg cttctgcccc
     3961 taacaaacat ggccgcccac tctgtcccac cccgcccttt ggctggtatc aaatactaga
     4021 ctacctttca tttccgggtc actgtagatg cgatggcgcc gattccaaag actgtggggc
     4081 ggatcaagct aggtgagcgg tatgacaaga ccagtggggg tctaggaggg cagaggcagc
     4141 ccctcttggg tccacctgtg gtctgaacgt ttcgagtagg attttatgca gagggcggag
     4201 agtcgagggt agagtgggga ggactctagc ccatctgtcc cgagcccgct gacgaggctc
     4261 cgccttcctt tcctgtagac tgctctctac ggccctagat cggtaccatg gtgagcaagg
     4321 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     4381 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     4441 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     4501 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     4561 ccgactactt gaagctgtcc ttccccgagg gcttcaattg ggagcgcgtg atgaacttcg
     4621 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     4681 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagtgtcgta
     4741 ccatgggctg ggaggcctcc actgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     4801 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     4861 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcgac atcaagttgg
     4921 acatcctttc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     4981 gccactccac cggcggcatg gacgagctgt acaaggagct ctagagggcc cgtttaaacc
     5041 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     5101 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     5161 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     5221 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     5281 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     5341 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5401 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5461 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5521 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5581 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5641 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5701 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5761 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5821 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5881 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5941 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6001 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     6061 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     6121 ccattttcgg atctgatcag cacgtgatga aaaagcctga actcaccgcg acgtctgtcg
     6181 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     6241 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     6301 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     6361 tcccgattcc ggaagtgctt gacattgggg aatttagcga gagcctgacc tattgcatct
     6421 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     6481 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     6541 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     6601 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     6661 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     6721 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     6781 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     6841 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     6901 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     6961 accaactcta tcagagcttg g
//
