LOCUS       sequence-202172-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Name: pBluescript-Delta-RRE. Blue Heron Biotechnology Inc. 
            bidirectional mutant variant of pBluescript-LANApi,K14 (pDD2000), 
            RRE..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  AUTHORS   Hilton IB, Dittmer DP
  TITLE     Quantitative analysis of the bidirectional viral G-protein-coupled 
            receptor and lytic latency-associated nuclear antigen promoter of 
            Kaposi's sarcoma-associated herpesvirus.
  JOURNAL   J Virol. 2012 Sep;86(18):9683-95. doi: 10.1128/JVI.00881-12. Epub 
            2012 Jun 27.
  PUBMED    22740392
REFERENCE   2  (bases 1 to 7146)
  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"; doi: 
            "10.1128/JVI.00881-12"; journalName: "J Virol"; date: "2012-09"; 
            volume: "86"; issue: "18"; pages: "9683-95"
COMMENT     Sequence Label: pDD2038
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     75..91
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     75..91
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    202..323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(366..1994)
                     /codon_start=1
                     /product="red-emitting variant of click beetle luciferase"
                     /label=CBRluc
                     /note="human codon-optimized"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RYGTQLIPGVTVLVYLPFFHAFGFHITLGYFMVGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLVDKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSAIIQTLGDEFKSGSLGRVTPLMAAKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGTEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLVKAGG"
     RBS             1036..1044
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      1991..2000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2401..2410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2407..4035
                     /codon_start=1
                     /product="green-emitting variant of click beetle 
                     luciferase"
                     /label=CBG68luc
                     /note="human codon-optimized"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RVGTQLIPGVTVLVYLPFFHAFGFSITLGYFMVGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLVDKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSANIHSLRDEFKSGSLGRVTPLMAAKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGKEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLEKAGG"
     RBS             3882..3890
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4092..4213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4359..4377)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4398..4414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4398..4414)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4411..4433)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4422..4438
                     /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(4446..4452,4453..4470,4471..4476))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4491..4512
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4629..4646)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4800..5388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4880..4899)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5559..6350,6351..6419))
                     /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     6182..6201
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6420..6524)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6551..7006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6638..6657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6848..6869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(7132..7146,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7146,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gagctccacc
       61 gcggtggcgg ccgctctaga actagtggat cccccgatcc ttatcgattt taccacattt
      121 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
      181 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
      241 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      301 tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc ggccggccgc cccgactcta
      361 gattactaac cgccggcctt caccaacaat tgtttcaaca gctccttgcg ggtaattttg
      421 cctgttacgt tacgagggat ggagtcaaca aaacgcacgc cgccacgcaa gtacttagta
      481 tggctcacac gttcagccag gtaatcatac acttctttgg cggtaatttc tgtaccaggc
      541 tgcttgacaa cgaaagcaga aggcagttcg ccggcctcca gatcaggaat gccgaccaca
      601 gcgacatcgc gaatgcatgg atttttcaac agaatctcct ccaactcagc tggagcaacc
      661 tggctaccct tgtatttgat cagctccttg taacgatcca cgacgtaaaa atgctcatct
      721 tcgtcgtaat atccaaaatc accagaatgc aaccagccgt cgtcgtcgat ggcctccttg
      781 gtagcttcaa cgttattgac ataacccttg ctcaccatag ggcctttgat acacagctcg
      841 cccacttggt tcgggcccaa agccttacca gtttcgcgat cagcgatctt agcagccatg
      901 agtggagtga cacggcccaa agagccgctc ttaaactcat ccccgagagt ctggataatc
      961 gcactggtag attcggtgag gccgaagcca caacgaatcc ctggaagatt caagcgtttg
     1021 gcggccactt cagcgacctc cttagccagt ggagcggcac cgcaacacaa ttcacgcagt
     1081 gaagacaagt cgtacttgtc cacgagtggg ctcttagaca aaaacaggat cacgctaggc
     1141 acgttgatga cactgcggac ttcataatct tggatggctt tcaagaaagc ctcctgatca
     1201 aaacggcgga acataatcac gcggagaccg accataaagt aacccaaagt aatatgaaag
     1261 ccgaaagcat ggaagaaagg caagtagacc aagacggtga caccaggaat cagctgagtg
     1321 ccgtagcgtg gatcgagagc atggatcaga cgcacgcaaa tgttttgatg ggtctgcatg
     1381 actccctttg ggagtccagt agtaccgctg ctacacagaa tggctgcaac ttgttccaca
     1441 gggtcgaagt ggagtggttt aaagtttgcg atgttgccgt ctgaatagcg agagatgaaa
     1501 ttaggcaaag attcgcaacc gtgaatattc tccacagtgt ccaagatgat gatacgctta
     1561 ataaagttgg tgcggctttg gacttccagg actttgttca gaatattctt agtggtgaag
     1621 acaatctgtg gcttagagat acccatgact ttacacagtt cgtcgggaat gtagctctcg
     1681 ttgactggag ccacgatcat accgatatac catgcggcga tgactggaat gaagaaacgg
     1741 gtattgtttt cagcacagat actaacgacg tcgttcatct tgtagccaca attgtggagg
     1801 gactgagcca gcaagacggt tgcctcaaaa aactccttgt agctcaaaga ttcatcgccg
     1861 accacatcga ccaaggcttg aggcaaatga gagtgcttgc ggagagcacg aaacagcatt
     1921 tcgccggcag tcaaatcctc caaaggatgg agaggctcag ggccatagat gacatttttc
     1981 tcacgcttta ccatggtggc tttaccaaca gtaccggaat gccaagctta cttagatcgc
     2041 agatctcgag cccgggctgc aggaattcga taggcagctg cgccacgaag cagtcacgtc
     2101 cccaagagca gcagcttggt ccggctgact tataaacagt gagtagcgcc cgcccacttg
     2161 tttcgggcgg cccgtaaggc acccttatct ttgaaatgct ataagatgtg tgggaaatag
     2221 taataccagg ctactgtgat accagttaag gtgtgtggtg gttttcgaaa aacatgggtt
     2281 attggccgtt tctgtttctc acgcccggat tatatatctg gacttgccta ggtagcatcc
     2341 atggcaacgc aaggccagca aacccactat caagcttggc attccggtac tgttggtaaa
     2401 gccaccatgg tgaaacgcga aaagaacgtg atctacggcc cagaaccact gcatccactg
     2461 gaagacctca ccgctggtga gatgctcttc cgagcactgc gtaaacatag tcacctccct
     2521 caagcactcg tggacgtcgt gggagacgag agcctctcct acaaagaatt tttcgaagct
     2581 actgtgctgt tggcccaaag cctccataat tgtgggtaca aaatgaacga tgtggtgagc
     2641 atttgtgctg agaataacac tcgcttcttt attcctgtaa tcgctgcttg gtacatcggc
     2701 atgattgtcg cccctgtgaa tgaatcttac atcccagatg agctgtgtaa ggttatgggt
     2761 attagcaaac ctcaaatcgt ctttactacc aaaaacatct tgaataaggt cttggaagtc
     2821 cagtctcgta ctaacttcat caaacgcatc attattctgg ataccgtcga aaacatccac
     2881 ggctgtgaga gcctccctaa cttcatctct cgttacagcg atggtaatat cgctaatttc
     2941 aagcccttgc attttgatcc agtcgagcaa gtggccgcta ttttgtgctc ctccggcacc
     3001 actggtttgc ctaaaggtgt catgcagact caccagaata tctgtgtgcg tttgatccac
     3061 gctctcgacc ctcgtgtggg tactcaattg atccctggcg tgactgtgct ggtgtatctg
     3121 cctttctttc acgcctttgg tttctctatt accctgggct atttcatggt cggcttgcgt
     3181 gtcatcatgt ttcgtcgctt cgaccaagaa gccttcttga aggctattca agactacgag
     3241 gtgcgttccg tgatcaacgt cccttcagtc attttgttcc tgagcaaatc tcctttggtt
     3301 gacaagtatg atctgagcag cttgcgtgag ctgtgctgtg gcgctgctcc tttggccaaa
     3361 gaagtggccg aggtcgctgc taagcgtctg aacctccctg gtatccgctg cggttttggt
     3421 ttgactgaga gcacttctgc taacatccat agcttgcgag acgagtttaa gtctggtagc
     3481 ctgggtcgcg tgactcctct tatggctgca aagatcgccg accgtgagac cggcaaagca
     3541 ctgggcccaa atcaagtcgg tgaattgtgt attaagggcc ctatggtctc taaaggctac
     3601 gtgaacaatg tggaggccac taaagaagcc attgatgatg atggctggct ccatagcggc
     3661 gacttcggtt actatgatga ggacgaacac ttctatgtgg tcgatcgcta caaagaattg
     3721 attaagtaca aaggctctca agtcgcacca gccgaactgg aagaaatttt gctgaagaac
     3781 ccttgtatcc gcgacgtggc cgtcgtgggt atcccagact tggaagctgg cgagttgcct
     3841 agcgcctttg tggtgaaaca acccggcaag gagatcactg ctaaggaggt ctacgactat
     3901 ttggccgagc gcgtgtctca caccaaatat ctgcgtggcg gcgtccgctt cgtcgattct
     3961 attccacgca acgttaccgg taagatcact cgtaaagagt tgctgaagca actcctcgaa
     4021 aaagctggcg gctagtaaag tcttcatgat tatctagagt cggggcggcc ggccgcttcg
     4081 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4141 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4201 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     4261 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     4321 gtcgacctcg agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg
     4381 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4441 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4501 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4561 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4621 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4681 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4741 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4801 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4861 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4921 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4981 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5041 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5101 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5161 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5221 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5281 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5341 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5401 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5461 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5521 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5581 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5641 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5701 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5761 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5821 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5881 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5941 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6001 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6061 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6121 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6181 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6241 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6301 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6361 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6421 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6481 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6541 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6601 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6661 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6721 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6781 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6841 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6901 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6961 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     7021 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     7081 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     7141 gacgtt
//
