LOCUS       sequence-367108-        8234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  K10.5 (vIRF3) tagged.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8234)
  TITLE     Dittmer lab plasmids Jan 2022
REFERENCE   2  (bases 1 to 8234)
  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"
COMMENT     Sequence Label: pOME0335
FEATURES             Location/Qualifiers
     source          1..8234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    914..938
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      977..986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2681..2746
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     2856..2875
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3047..3071)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3082..3084,3085..3087,3088..3801)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(3115..3143)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3738..3758
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3823..3840)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3829..4053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4099..4527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4186..4205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4396..4417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4536..4556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4541..4870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4721..4856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4783..4802
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4937..5731
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4991..5010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5601..5620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5907..6028
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5944..5963)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5998..6017
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6077..6093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6077..6093)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6090..6112)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6101..6117
                     /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(6125..6131,6132..6149,6150..6155))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6170..6191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6308..6325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6479..7067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6559..6578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7238..8029,8030..8098))
                     /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     7861..7880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8099..8203)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttgat caaacaagtt tgtacaaaaa agcaggctct ttaaaggaac caattcagtc
      961 gactggatcc ggtaccgcca ccatggcggg acgcaggctt acctggattt ctgagtttat
     1021 tgtaggtgct ttggactctg ataaatatcc tttggtcaag tggctagata gatctactgg
     1081 aacatttctt gctccggctg cccgtaatga cgtaattcct ctggatagcc tacagttttt
     1141 cattgatttt aagagggaat gcctatcgaa gggcctgcat cccagagatt tactgggctc
     1201 gccgattacg gcttttggga aaatatgtac cacgtcgcgg cgccttagac gcttgccagg
     1261 tgaagagtac gaggtcgtac agggaattaa ttgtagaagg tggcgcctcc tgtgtgccga
     1321 ggtaaaggaa tgctggtggt gcgttcatgc caggactcac ctacacagtg ggtcatcact
     1381 atgggaaatt ttgtatcaac acagtgtacg gctcgagaag catcggagaa gaccaaggcc
     1441 atttgtgggt gaaaactcgg attccagtga ggaggatcac ccagcctttt gcgatgtgcc
     1501 ggtcacgcag acgggcgcgg aatctgagga ctctggagac gagggaccat cgacgcgcca
     1561 tagtgcgtct ggggttcagc cagttgatga tgccaatgcc gactctcctg gctctggaga
     1621 cgaaggaccc tcgacgcgtc atagcgactc gcagcccccc ccggccaatg aaacaacggt
     1681 gcacacagac aacgttgaag atgacctcac actgcttgat aaagaatctg catgtgcatt
     1741 gatgtaccac gtgggacagg agatggacat gctaatgagg gcgatgtgcg atgaagacct
     1801 ctttgatctg cttggcatcc cagaggatgt tatcgcaaca tcacagcccg gaggcgacac
     1861 ggatgcaagc ggcgtggtaa cagagggctc aatcgccgcc tcggctgtcg gggcgggtgt
     1921 agaggatgtg tacttagctg gggcactcga ggcccagaat gtagcagggg aatatgtgtt
     1981 ggagataagt gacgaagaag tcgatgatgg tgctggactg ccgccggcgt ccagacgccg
     2041 gccagttgtt ggcgaatttt tatgggatga tgggccacgg agacacgaga ggcctaccac
     2101 gaggcgcatt cgccacagga agcttagatc cgcatattat agagtggcac ggccgccagt
     2161 aatgataacc gataggcttg gtgtggaagt gttttatttt ggccgccctg caatgtcttt
     2221 ggaagtggaa cgaaaggtgt ttattctatg ttcccagaac ccactggcag acattagcca
     2281 ctcttgcttg cattcgcgca aagggttaag agttttgttg cccaaacctg acgacaataa
     2341 cacagggcca ggagacgtta acctgctggc ggccgtgctg cgctcgtttg cttcgggtct
     2401 tgtgatagtt tctctccgat ctggcattta tgttaagaat ttgtgcaagt ctaccgtatt
     2461 atatcatgga aataatcctc caaagaagtt tggtgtgatc tgcggactgt catctagggc
     2521 tgttctggat gtttttaatg tggcacaata tcgcatacag ggacatgagc acattaaaaa
     2581 aacaactgtg ttcatcggag gtgacccaac gtcggcagaa cagttcgata tggtccccct
     2641 cgtcatcaag ctcagattgc gttcagttac atgtgatgac gactacaaag accatgacgg
     2701 tgattataaa gatcatgata tcgattacaa ggatgacgat gacaagtagt aacccgggcg
     2761 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtcc cccgtgaact
     2821 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     2881 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     2941 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
     3001 ccagtctgca ggtcgacgaa ttcgcggccg cactcgagat atctagaccc agctttcttg
     3061 tacaaagtgg ttcgatctag aatggctagc aaaggagaag aacttttcac tggagttgtc
     3121 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     3181 ggtgaaggtg atgctacata cggaaagctt acccttaaat ttatttgcac tactggaaaa
     3241 ctacctgttc catggccaac acttgtcact actttctctt atggtgttca atgcttttcc
     3301 cgttatccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     3361 gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc
     3421 aagtttgaag gtgataccct tgttaatcgt atcgagttaa aaggtattga ttttaaagaa
     3481 gatggaaaca ttctcggaca caaactcgag tacaactata actcacacaa tgtatacatc
     3541 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa
     3601 gatggatccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     3661 gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac
     3721 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
     3781 atggatgagc tctacaaata atgaattaaa cccgctgatc agcctcgact gtgccttcta
     3841 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3901 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3961 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4021 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4081 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4441 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4561 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4621 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4681 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4741 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4801 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4861 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     4921 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4981 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5041 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5101 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5161 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5221 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5281 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5341 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5401 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5461 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5521 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5581 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5641 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5701 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     5761 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5821 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5881 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5941 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6001 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6061 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6121 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6181 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6241 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6301 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6361 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6421 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6481 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6541 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6601 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6661 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6721 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6781 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6841 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6901 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6961 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7021 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7081 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7141 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7201 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7261 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7321 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7381 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7441 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7501 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7561 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7621 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7681 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7741 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7801 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7861 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7921 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7981 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8041 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8101 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8161 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8221 tgccacctga cgtc
//
