LOCUS       sequence-194117-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Name: pBluescript-RBPjk-Delta-B. RBPjk binding site B mutation in 
            pBluescript-LANApi,K14 (pDD2000)..
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: pDD2030
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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)."
     primer_bind     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        224..242
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    388..509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(566..2194)
                     /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
                     PCIRDVAAVGIPDLEAGELPSAFVVKQPGKEITAKEVYDYLAERVSHTKYLCGGVRFVD
                     SIPRNVTGKITRKELLKQLLEKAGG"
     RBS             711..719
                     /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      2191..2200
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2601..2610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2607..4235
                     /codon_start=1
                     /product="red-emitting variant of click beetle luciferase"
                     /label=CBRluc
                     /note="human codon-optimized"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RYGTQLIPGVTVLVYLPFFHAFGFHITLGYFMVGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLADKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSAIIQTLGDEFKSGSLGRVTPLMAAKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGTEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLVKAGG"
     RBS             3557..3565
                     /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    4278..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4510..4526)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4510..4526)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4558..4577)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4559..4577)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4584..4601)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4584..4600)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4593..4615)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(4741..5196)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4878..4899)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5090..5109
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5223..5327
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5328..5396,5397..6188)
                     /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     complement(5546..5565)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6359..6947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6848..6867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7101..7118
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
      241 gggaacaaaa gctggtaccg ggccccccct cgaggtcgac ggatccttat cgattttacc
      301 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
      361 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
      421 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
      481 ggtttgtcca aactcatcaa tgtatcttat catgtctgct cgaagcggcc ggccgccccg
      541 actctagata atcatgaaga ctttactagc cgccagcttt ttcgaggagt tgcttcagca
      601 actctttacg agtgatctta ccggtaacgt tgcgtggaat agaatcgacg aagcggacgc
      661 cgccacacag atatttggtg tgagacacgc gctcggccaa atagtcgtag acctccttag
      721 cagtgatctc cttgccgggt tgtttcacca caaaggcgct aggcaactcg ccagcttcca
      781 agtctgggat acccacggcg gccacgtcgc ggatacaagg gttcttcagc aaaatttctt
      841 ccagttcggc tggtgcgact tgagagcctt tgtacttaat caattctttg tagcgatcga
      901 ccacatagaa gtgttcgtcc tcatcatagt aaccgaagtc gccgctatgg agccagccat
      961 catcatcaat ggcttcttta gtggcctcca cattgttcac gtagccttta gagaccatag
     1021 ggcccttaat acacaattca ccgacttgat ttgggcccag tgctttgccg gtctcacggt
     1081 cggcgatctt tgcagccata agaggagtca cgcgacccag gctaccagac ttaaactcgt
     1141 ctcgcaagct atggatgtta gcagaagtgc tctcagtcaa accaaaaccg cagcggatac
     1201 cagggaggtt cagacgctta gcagcgacct cggccacttc tttggccaaa ggagcagcgc
     1261 cacagcacag ctcacgcaag ctgctcagat catacttgtc aaccaaagga gatttgctca
     1321 ggaacaaaat gactgaaggg acgttgatca cggaacgcac ctcgtagtct tgaatagcct
     1381 tcaagaaggc ttcttggtcg aagcgacgaa acatgatgac acgcaagccg accatgaaat
     1441 agcccagggt aatagagaaa ccaaaggcgt gaaagaaagg cagatacacc agcacagtca
     1501 cgccagggat caattgagta cccacacgag ggtcgagagc gtggatcaaa cgcacacaga
     1561 tattctggtg agtctgcatg acacctttag gcaaaccagt ggtgccggag gagcacaaaa
     1621 tagcggccac ttgctcgact ggatcaaaat gcaagggctt gaaattagcg atattaccat
     1681 cgctgtaacg agagatgaag ttagggaggc tctcacagcc gtggatgttc tcgacggtat
     1741 ccagaataat gatgcgtttg atgaagttag tacgagactg gacttccaag accttattca
     1801 agatgttttt ggtagtaaag acgatttgag gtttgctaat acccataacc ttacacagct
     1861 catctgggat gtaagattca ttcacagggg cgacaatcat gccgatgtac caagcagcga
     1921 ttacaggaat aaagaagcga gtgttattct cagcacaaat gctcaccaca tcgttcattt
     1981 tgtacccaca attatggagg ctttgggcca acagcacagt agcttcgaaa aattctttgt
     2041 aggagaggct ctcgtctccc acgacgtcca cgagtgcttg agggaggtga ctatgtttac
     2101 gcagtgctcg gaagagcatc tcaccagcgg tgaggtcttc cagtggatgc agtggttctg
     2161 ggccgtagat cacgttcttt tcgcgtttca ccatggtggc tttaccaaca gtaccggaat
     2221 gccaagcttg atagtgggtt tgctggcctt gcgttgccat ggatgctacc taggcaagtc
     2281 cagatatata atccgggcgt gagaaacaga aacggccaat aacccatgtt tttcgaaaac
     2341 caccacacac cttaacacaa atcatgtaca cctggtatta ctatgattac cacatcttat
     2401 agcatttcaa agataagggt gccttacggg ccgcccgaaa caagtgggcg ggcgctactc
     2461 actgtttata agtcagccgg accaagctgc tgctcttggg gacgtgactg cttcgtggcg
     2521 cagctgccta tcgaattcct gcagcccggg ctcgagatct gcgatctaag taagcttggc
     2581 attccggtac tgttggtaaa gccaccatgg taaagcgtga gaaaaatgtc atctatggcc
     2641 ctgagcctct ccatcctttg gaggatttga ctgccggcga aatgctgttt cgtgctctcc
     2701 gcaagcactc tcatttgcct caagccttgg tcgatgtggt cggcgatgaa tctttgagct
     2761 acaaggagtt ttttgaggca accgtcttgc tggctcagtc cctccacaat tgtggctaca
     2821 agatgaacga cgtcgttagt atctgtgctg aaaacaatac ccgtttcttc attccagtca
     2881 tcgccgcatg gtatatcggt atgatcgtgg ctccagtcaa cgagagctac attcccgacg
     2941 aactgtgtaa agtcatgggt atctctaagc cacagattgt cttcaccact aagaatattc
     3001 tgaacaaagt cctggaagtc caaagccgca ccaactttat taagcgtatc atcatcttgg
     3061 acactgtgga gaatattcac ggttgcgaat ctttgcctaa tttcatctct cgctattcag
     3121 acggcaacat cgcaaacttt aaaccactcc acttcgaccc tgtggaacaa gttgcggcca
     3181 ttctgtgtag cagcggtact actggactcc caaagggagt catgcagacc catcaaaaca
     3241 tttgcgtgcg tctgatccat gctctcgatc cacgctacgg cactcagctg attcctggtg
     3301 tcaccgtctt ggtctacttg cctttcttcc atgctttcgg ctttcatatt actttgggtt
     3361 actttatggt cggtctccgc gtgattatgt tccgccgttt tgatcaggag gctttcttga
     3421 aagccatcca agattatgaa gtccgcagtg tcatcaacgt gcctagcgtg atcctgtttt
     3481 tgtctaagag cccactcgcg gacaagtacg acttgtcttc actgcgtgaa ttgtgttgcg
     3541 gtgccgctcc actggctaag gaggtcgctg aagtggccgc caaacgcttg aatcttccag
     3601 ggattcgttg tggcttcggc ctcaccgaat ctaccagtgc gattatccag actctcgggg
     3661 atgagtttaa gagcggctct ttgggccgtg tcactccact catggctgct aagatcgctg
     3721 atcgcgaaac tggtaaggct ttgggcccga accaagtggg cgagctgtgt atcaaaggcc
     3781 ctatggtgag caagggttat gtcaataacg ttgaagctac caaggaggcc atcgacgacg
     3841 acggctggtt gcattctggt gattttggat attacgacga agatgagcat ttttacgtcg
     3901 tggatcgtta caaggagctg atcaaataca agggtagcca ggttgctcca gctgagttgg
     3961 aggagattct gttgaaaaat ccatgcattc gcgatgtcgc tgtggtcggc attcctgatc
     4021 tggaggccgg cgaactgcct tctgctttcg ttgtcaagca gcctggtaca gaaattaccg
     4081 ccaaagaagt gtatgattac ctggctgaac gtgtgagcca tactaagtac ttgcgtggcg
     4141 gcgtgcgttt tgttgactcc atccctcgta acgtaacagg caaaattacc cgcaaggagc
     4201 tgttgaaaca attgttggtg aaggccggcg gttagtaatc tagagtcggg gcggccggcc
     4261 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     4321 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     4381 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     4441 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     4501 ggatcggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc caattcgccc
     4561 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4621 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4681 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4741 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4801 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaggaa tagaccgaga
     4861 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4921 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4981 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     5041 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     5101 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     5161 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     5221 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5281 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5341 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5401 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5461 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5521 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5581 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5641 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5701 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5761 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5821 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5881 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5941 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6001 ctggctggtt tattgctgat aaatctggag ccggtgagcg cgggtctcgc ggtatcattg
     6061 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6121 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6181 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6241 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6301 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6361 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6421 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6481 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6541 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6601 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6661 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6721 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6781 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6841 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6901 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6961 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     7021 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     7081 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaggg cgcccaatac
     7141 gcaaac
//
