LOCUS       sequence-194107-        5128 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Name: pCBR-RBPjk-Delta-A. RBPjk binding site A mutation in pCBR-K14 
            (pDD2005)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5128)
  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 5128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pDD2028
FEATURES             Location/Qualifiers
     source          1..5128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      397..406
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             403..2031
                     /codon_start=1
                     /product="red-emitting variant of click beetle luciferase"
                     /label=CBRluc
                     /note="human codon-optimized"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RYGTQLIPGVTVLVYLPFFHAFGFHITLGYFMVGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLVDKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSAIIQTLGDEFKSGSLGRVTPLMATKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGTEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLVKAGG"
     RBS             1353..1361
                     /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    2074..2195
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2085..2104)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2139..2158
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2443..2460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2614..3202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2694..2713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3373..4164,4165..4233))
                     /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     3996..4015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4234..4338)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4365..4820
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4452..4471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4662..4683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4951..4999
                     /note="synthetic polyadenylation signal"
     misc_feature    5013..5104
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5053..5072
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     join(5128,1..16)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 cgaggtcgac ggtatcgata agcttgatag tgggtttgct ggccttgcgt tgccatggat
       61 gctacctagg caagtccaga tatataatcc gggcgtatac aacagaaacg gccaataacc
      121 catgtttttc gaaaaccacc acacacctta acacaaatca tgtacacctg gtattactat
      181 ttcccacaca tcttatagca tttcaaagat aagggtgcct tacgggccgc ccgaaacaag
      241 tgggcgggcg ctactcactg tttataagtc agccggacca agctgctgct cttggggacg
      301 tgactgcttc gtggcgcagc tgcctatcga attcctgcag cccgggctcg agatctgcga
      361 tctaagtaag cttggcattc cggtactgtt ggtaaagcca ccatggtaaa gcgtgagaaa
      421 aatgtcatct atggccctga gcctctccat cctttggagg atttgactgc cggcgaaatg
      481 ctgtttcgtg ctctccgcaa gcactctcat ttgcctcaag ccttggtcga tgtggtcggc
      541 gatgaatctt tgagctacaa ggagtttttt gaggcaaccg tcttgctggc tcagtccctc
      601 cacaattgtg gctacaagat gaacgacgtc gttagtatct gtgctgaaaa caatacccgt
      661 ttcttcattc cagtcatcgc cgcatggtat atcggtatga tcgtggctcc agtcaacgag
      721 agctacattc ccgacgaact gtgtaaagtc atgggtatct ctaagccaca gattgtcttc
      781 accactaaga atattctgaa caaagtcctg gaagtccaaa gccgcaccaa ctttattaag
      841 cgtatcatca tcttggacac tgtggagaat attcacggtt gcgaatcttt gcctaatttc
      901 atctctcgct attcagacgg caacatcgca aactttaaac cactccactt cgaccctgtg
      961 gaacaagttg cagccattct gtgtagcagc ggtactactg gactcccaaa gggagtcatg
     1021 cagacccatc aaaacatttg cgtgcgtctg atccatgctc tcgatccacg ctacggcact
     1081 cagctgattc ctggtgtcac cgtcttggtc tacttgcctt tcttccatgc tttcggcttt
     1141 catattactt tgggttactt tatggtcggt ctccgcgtga ttatgttccg ccgttttgat
     1201 caggaggctt tcttgaaagc catccaagat tatgaagtcc gcagtgtcat caacgtgcct
     1261 agcgtgatcc tgtttttgtc taagagccca ctcgtggaca agtacgactt gtcttcactg
     1321 cgtgaattgt gttgcggtgc cgctccactg gctaaggagg tcgctgaagt ggccgccaaa
     1381 cgcttgaatc ttccagggat tcgttgtggc ttcggcctca ccgaatctac cagtgcgatt
     1441 atccagactc tcggggatga gtttaagagc ggctctttgg gccgtgtcac tccactcatg
     1501 gctactaaga tcgctgatcg cgaaactggt aaggctttgg gcccgaacca agtgggcgag
     1561 ctgtgtatca aaggccctat ggtgagcaag ggttatgtca ataacgttga agctaccaag
     1621 gaggccatcg acgacgacgg ctggttgcat tctggtgatt ttggatatta cgacgaagat
     1681 gagcattttt acgtcgtgga tcgttacaag gagctgatca aatacaaggg tagccaggtt
     1741 gctccagctg agttggagga gattctgttg aaaaatccat gcattcgcga tgtcgctgtg
     1801 gtcggcattc ctgatctgga ggccggcgaa ctgccttctg ctttcgttgt caagcagcct
     1861 ggtacagaaa ttaccgccaa agaagtgtat gattacctgg ctgaacgtgt gagccatact
     1921 aagtacttgc gtggcggcgt gcgttttgtt gactccatcc ctcgtaacgt aacaggcaaa
     1981 attacccgca aggagctgtt gaaacaattg ttggtgaagg ccggcggtta gtaatctaga
     2041 gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     2101 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     2161 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     2221 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     2281 gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     2341 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     2401 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     2461 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2521 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2581 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2641 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2701 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2761 cggatacctg tccgcctttc tcccttcggg aggcgtggcg ctttctcata gctcacgctg
     2821 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2881 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2941 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3001 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3061 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3121 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3181 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3301 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatmt atgagtaaac
     3361 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3421 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3481 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3541 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3601 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3661 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3721 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3781 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3841 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3901 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3961 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4021 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4081 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4141 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4201 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4261 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4321 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     4381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4621 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4681 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4741 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4801 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     4861 atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat
     4921 taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg
     4981 tgtgttggtt ttttgtgtga atcgatagta ctaacatacg ctctccatca aaacaaaacg
     5041 aaacaaaaca aactagcaaa ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt
     5101 ctctatcgat aggtaccggg ccccccct
//
