LOCUS       sequence-193508-        6875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mVirD2-GFP expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6875)
  AUTHORS   Bernardinelli G, Hogberg B
  TITLE     Entirely enzymatic nanofabrication of DNA-protein conjugates.
  JOURNAL   Nucleic Acids Res. 2017 Oct 13;45(18):e160. doi: 10.1093/nar/gkx707.
  PUBMED    28977490
REFERENCE   2  (bases 1 to 6875)
  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: "Nucleic 
            Acids Res."; date: "2017-10-13"; volume: "45(18):e160. doi"; pages: 
            " 10.1093/nar/gkx707"
COMMENT     Sequence Label: pmVirD2-GFP
FEATURES             Location/Qualifiers
     source          1..6875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      52..138
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     273..292
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        273..291
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    292..316
                     /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)."
     RBS             331..353
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        join(402..416,417..422,423..439,440..446)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    422..438
                     /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)."
     protein_bind    454..470
                     /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)."
     RBS             489..500
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             510..527
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(1170..1172,1173..1886)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1212..1233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1473..1492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1820..1841
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1889..1920
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     complement(1974..1992)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1988..2035
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2095..2199
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2200..2268,2269..3060)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2418..2437)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3102..3615)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3509..3528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3726..4314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4215..4234
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4468..4485
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4507..4526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4667..4689
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4683..4874)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5431..6513)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6475..6494
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6514..6591)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      6744..6830
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tggcgggcag gacgcccgcc ataaactgcc aggaattaat tccccaggca tcaaataaaa
       61 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      121 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
      181 gggtggcggg caggacgccc gccataaact gccaggaatt ggggatcgga attaattccc
      241 ggtttaaacc ggggatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
      301 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
      361 atggctagca ccatggatat caagcttacc ggtgaattca atcataaaaa atttatttgc
      421 tttgtgagcg gataacaatt ataatagatt caattgtgag cggataacaa tttcacacag
      481 atatcattaa agaggagaaa tctagaatgc atcatcatca tcatcatccc gatcgcgctc
      541 aagtaatcat tcgcattgtg ccaggaggtg gaaccaagac ccttcagcag ataatcaatc
      601 agttggagta cctgtcccgt aagggaaagc tggaactgca gcgttcagcc cggcatctcg
      661 atattcccgt tccgccggat caaatccgtg agcttgccca aagctgggtt acggaggccg
      721 ggatttatga cgaaagtcag tcagacgatg ataggcaaca agacttaaca acacacatta
      781 ttgtaagctt ccccgcaggt accgaccaaa ccgcagctta tgaagccagc cgggaatggg
      841 cagccgagat gtttgggtca ggatacgggg gtggccgcta taactatctg acagcctacc
      901 acgtcgaccg cgatcatcca catttacatg tcgtggtcaa tcgtcgggaa cttctggggc
      961 acgggtggct gaaaatatcc aggcgccatc cccagctgaa ttatgacggc ttacggaaaa
     1021 agatggcaga gatttcactt cgtcacggca tagtcctgga tgcgacttcg cgagcagaaa
     1081 ggggaatagc agagcgacca atcacatatg ctgaacatcg ccgccttgag cggatgcagc
     1141 catggggtgg ttctggtggt ggttctggtg tgagcaaggg cgaggagctg ttcaccgggg
     1201 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1261 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1321 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cacctacggc gtgcagtgct
     1381 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1441 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1501 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1561 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaaggtct
     1621 atatcaccgc cgacaagcag aagaacggca tcaaggtgaa cttcaagacc cgccacaaca
     1681 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1741 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1801 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1861 tcggcatgga cgagctgtac aagtaaaaag tcaaaagcct ccgaccggag gcttttgact
     1921 tattgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1981 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     2041 aggaggaact atatccggat aactacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     2101 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2161 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2221 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2281 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2341 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg
     2401 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2461 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2521 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2581 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2641 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2701 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2761 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2821 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2881 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2941 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3001 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3061 ctgtcagacc aagtttactc atatatactt tagattgatt taccccggtt gataatcaga
     3121 aaagccccaa aaacaggaag attgtataag caaatattta aattgtaaac gttaatattt
     3181 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3241 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag
     3301 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3361 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     3421 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3481 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3541 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3601 cgctacaggg cgcgtaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3661 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3721 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3781 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3841 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     3901 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3961 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4021 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4081 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4141 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4201 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4261 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4321 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4381 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     4441 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc tatggtgcac
     4501 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4561 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4621 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4681 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4741 gcgtggtcgt gcagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4801 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     4861 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     4921 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     4981 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     5041 actcagggtc aatgccagcc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     5101 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5161 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5221 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5281 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5341 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     5401 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5461 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5521 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     5581 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     5641 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     5701 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     5761 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     5821 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     5881 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     5941 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     6001 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     6061 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     6121 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     6181 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     6241 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     6301 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     6361 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     6421 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     6481 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     6541 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     6601 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     6661 tgagcaccgc cgccgcaagg aatggtgcat gccggcatgc cgccctttcg tcttcaagaa
     6721 ttaattccca attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     6781 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     6841 cggatttgaa cgttgcgaag caacggcccg gaggg
//
