LOCUS       sequence-359393-        7731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KSHV K15 TAGGED.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7731)
  TITLE     Dittmer lab plasmids Jan 2022
REFERENCE   2  (bases 1 to 7731)
  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: pOME0288
FEATURES             Location/Qualifiers
     source          1..7731
                     /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"
     CDS             2443..2508
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2544..2568)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2579..2581,2582..2584,2585..3298)
                     /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(2612..2640)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3235..3255
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3320..3337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3326..3550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3596..4024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3683..3702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3893..3914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4033..4053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4038..4367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4218..4353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4280..4299
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4434..5228
                     /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(4488..4507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5098..5117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5404..5525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5441..5460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5495..5514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5574..5590)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5574..5590)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5587..5609)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5598..5614
                     /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(5622..5628,5629..5646,5647..5652))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5667..5688
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5805..5822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5976..6564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6056..6075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6735..7526,7527..7595))
                     /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     7358..7377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7596..7700)
                     /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 gacggtaccg ccaccatgaa gacactcata ttcttctgga atttatggct ttgggcccta
     1021 ctggtatgtt tttggtgtat cactcttgtc tgtgtaacta ccaactcaat tgatacaatg
     1081 gcttctttgc ttgttatgtg cattttgttt gtcagtgcta ttaataagta tacacaggca
     1141 atctctagca acaatcctaa atggccttca tcctggcacc taggaattat tgcttgcatt
     1201 gtccttaaac tttggaactt gtcaactacc aactctgtaa cctatgcttg cttaatcacc
     1261 acggccattt tatcactggt aacagctttt ttgacgttaa taaaacactg caccgcctgt
     1321 aaattacaac ttgaacatgg aatattattc acatcaacgt ttgctgtgtt gatgacaaac
     1381 atgctggtcc acatgtcaaa cacctggcaa tcgtcttgga tattttttcc aattagcttc
     1441 actctcagct tgccattctt gtatgcattt gccaccgtga aaacaggcaa tattaaatta
     1501 gtgtcatctg tctctttcat ttgtgcaggc ttggtcatgg gttaccctgt atcctgctgt
     1561 aaaacgcata catgtactgc caccgctgca ggattaagtc tttccagcat ttatttggga
     1621 ttcactggca taatcagcac tcttcataag tcatgggctc caccaaaacg tggaatatta
     1681 acgttcctgc ttcttcaggg gggagtgcta accacacaaa ctttaacaac cgaactactt
     1741 gcaattacta gcaccactgg caatatcaaa ggccacgaaa tattgcttct tgtatgcctc
     1801 atttttctat ggtgccttta tgtgtggcaa agttttaaca aggcgtctct tgttactgga
     1861 atgctgcatt taattgcagc gtggtcccac actggaggtt gtgtacaatt agtaatgctt
     1921 ctccctagtg gtttaacaag aggcatccta acaatgatca tctgcatcag tacactattc
     1981 agtacgttac aaggactgct ggtattttat ttatataagg aaaaaaaagt ggtggctgta
     2041 aatagttacc gacagagaag gcggcgcata tacacgcggg accagaactt acaccacaat
     2101 gacaaccacc ttggcaataa tgtaattagc cctccaccat taccaccttt ttttagacag
     2161 cctgttaggt tacctagcca tgtcaccgat aggggccgcg ggagccaacc actgaacgag
     2221 gtggagttac aggaagtgaa tagggaccca cccaatgtat tcgggtatgc cagtatttta
     2281 gtgtccggtg cggaagaatc acgtgaacca tcaccacagc cagaccaatc aggaatgtca
     2341 attttaaggg ttgatggtgg gagtgccttc cgtatagaca ccgcccaagc cgccacccaa
     2401 ccgacagacg acctgtatga ggaggtttta tttcccagga acgactacaa agaccatgac
     2461 ggtgattata aagatcatga tatcgattac aaggatgacg atgacaagta gtaactcgag
     2521 gcggccgcac tcgagatatc tagacccagc tttcttgtac aaagtggttc gatctagaat
     2581 ggctagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     2641 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg ctacatacgg
     2701 aaagcttacc cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact
     2761 tgtcactact ttctcttatg gtgttcaatg cttttcccgt tatccggatc atatgaaacg
     2821 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt
     2881 caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt
     2941 taatcgtatc gagttaaaag gtattgattt taaagaagat ggaaacattc tcggacacaa
     3001 actcgagtac aactataact cacacaatgt atacatcacg gcagacaaac aaaagaatgg
     3061 aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggatccgttc aactagcaga
     3121 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     3181 cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct
     3241 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct acaaataatg
     3301 aattaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3361 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3421 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3481 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3541 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4021 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4201 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4321 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4381 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4441 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4501 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4561 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4621 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4681 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4741 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4801 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     4861 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     4921 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     4981 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5041 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5101 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5161 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5221 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     5281 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5341 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5401 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5461 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5521 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5581 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5641 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5701 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5761 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
