LOCUS       sequence-193506-        6171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mVirD2 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6171)
  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 6171)
  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
FEATURES             Location/Qualifiers
     source          1..6171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      105..191
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     326..345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        326..344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    345..369
                     /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             384..406
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        join(455..469,470..475,476..492,493..499)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    475..491
                     /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    507..523
                     /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             542..553
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             563..580
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1323..1341)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1337..1384
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1444..1548
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1549..1617,1618..2409)
                     /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(1767..1786)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2451..2964)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2646..2667)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2858..2877
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3075..3663
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3564..3583
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3817..3834
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3856..3875)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4016..4038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4032..4223)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4780..5862)
                     /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     5824..5843
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5863..5940)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      join(6093..6171,1..8)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
       61 caggacgccc gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg
      121 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
      181 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
      241 gggcaggacg cccgccataa actgccagga attggggatc ggaattaatt cccggtttaa
      301 accggggatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
      361 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatggcta
      421 gcaccatgga tatcaagctt accggtgaat tcaatcataa aaaatttatt tgctttgtga
      481 gcggataaca attataatag attcaattgt gagcggataa caatttcaca cagatatcat
      541 taaagaggag aaatctagaa tgcatcatca tcatcatcat cccgatcgcg ctcaagtaat
      601 cattcgcatt gtgccaggag gtggaaccaa gacccttcag cagataatca atcagttgga
      661 gtacctgtcc cgtaagggaa agctggaact gcagcgttca gcccggcatc tcgatattcc
      721 cgttccgccg gatcaaatcc gtgagcttgc ccaaagctgg gttacggagg ccgggattta
      781 tgacgaaagt cagtcagacg atgataggca acaagactta acaacacaca ttattgtaag
      841 cttccccgca ggtaccgacc aaaccgcagc ttatgaagcc agccgggaat gggcagccga
      901 gatgtttggg tcaggatacg ggggtggccg ctataactat ctgacagcct accacgtcga
      961 ccgcgatcat ccacatttac atgtcgtggt caatcgtcgg gaacttctgg ggcacgggtg
     1021 gctgaaaata tccaggcgcc atccccagct gaattatgac ggcttacgga aaaagatggc
     1081 agagatttca cttcgtcacg gcatagtcct ggatgcgact tcgcgagcag aaaggggaat
     1141 agcagagcga ccaatcacat atgctgaaca tcgccgcctt gagcggatgc agccatggta
     1201 acgcaaaaaa ccccgcttcg gcggggtttt ttcgcggatc ccccgggctc gaggttaatt
     1261 aagcggccgc attgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     1321 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     1381 tttgctgaaa ggaggaacta tatccggata actacgtcag gtggcacttt tcggggaaat
     1441 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1501 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1561 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1621 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1681 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttc
     1741 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     1801 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1861 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1921 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1981 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2041 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2101 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2161 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2221 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2281 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2341 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2401 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt accccggttg
     2461 ataatcagaa aagccccaaa aacaggaaga ttgtataagc aaatatttaa attgtaaacg
     2521 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2581 aggccgaaat cggcaaaatc ccttataaat caaaagaata gcccgagata gggttgagtg
     2641 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2701 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcacccaaa tcaagttttt
     2761 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2821 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2881 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2941 ttaatgcgcc gctacagggc gcgtaaaagg atctaggtga agatcctttt tgataatctc
     3001 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3061 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3121 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3181 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     3241 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     3301 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3361 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3421 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3481 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3541 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3601 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3661 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3721 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3781 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagct
     3841 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3901 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3961 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     4021 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     4081 agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc cgcgtccagc
     4141 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     4201 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     4261 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     4321 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     4381 agaaaaatca ctcagggtca atgccagccg aacgccagca agacgtagcc cagcgcgtcg
     4441 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     4501 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     4561 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     4621 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     4681 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     4741 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4801 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4861 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     4921 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     4981 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     5041 gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg
     5101 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     5161 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     5221 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     5281 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     5341 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     5401 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     5461 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     5521 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     5581 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     5641 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     5701 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     5761 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     5821 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     5881 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     5941 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     6001 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg ccggcatgcc gccctttcgt
     6061 cttcaagaat taattcccaa ttccccaggc atcaaataaa acgaaaggct cagtcgaaag
     6121 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt a
//
