LOCUS       sequence-194526-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Name: pBluescript-LANApi,K14. Bidirectional reporter driving LANApi 
            side [green luciferase isoform from pCBG68] and K14 side [red 
            luciferase isoform from pCBR] in pBluescript backbone..
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: pDD2000
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(165..1793)
                     /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             835..843
                     /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      1790..1799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2200..2209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2206..3834
                     /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             3681..3689
                     /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    3891..4012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4158..4176)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4197..4213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4197..4213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4210..4232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4221..4237
                     /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(4245..4251,4252..4269,4270..4275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4290..4311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4428..4445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4599..5187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4679..4698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5358..6149,6150..6218))
                     /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     5981..6000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6219..6323)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6350..6805
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6437..6456)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6647..6668
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6931..6953
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6945..6962
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6946..6962
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6969..6988
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6969..6987
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7020..7036
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7020..7036
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct gctcgaagcg gccggccgcc ccgactctag attactaacc gccggccttc
      181 accaacaatt gtttcaacag ctccttgcgg gtaattttgc ctgttacgtt acgagggatg
      241 gagtcaacaa aacgcacgcc gccacgcaag tacttagtat ggctcacacg ttcagccagg
      301 taatcataca cttctttggc ggtaatttct gtaccaggct gcttgacaac gaaagcagaa
      361 ggcagttcgc cggcctccag atcaggaatg ccgaccacag cgacatcgcg aatgcatgga
      421 tttttcaaca gaatctcctc caactcagct ggagcaacct ggctaccctt gtatttgatc
      481 agctccttgt aacgatccac gacgtaaaaa tgctcatctt cgtcgtaata tccaaaatca
      541 ccagaatgca accagccgtc gtcgtcgatg gcctccttgg tagcttcaac gttattgaca
      601 taacccttgc tcaccatagg gcctttgata cacagctcgc ccacttggtt cgggcccaaa
      661 gccttaccag tttcgcgatc agcgatctta gcagccatga gtggagtgac acggcccaaa
      721 gagccgctct taaactcatc cccgagagtc tggataatcg cactggtaga ttcggtgagg
      781 ccgaagccac aacgaatccc tggaagattc aagcgtttgg cggccacttc agcgacctcc
      841 ttagccagtg gagcggcacc gcaacacaat tcacgcagtg aagacaagtc gtacttgtcc
      901 acgagtgggc tcttagacaa aaacaggatc acgctaggca cgttgatgac actgcggact
      961 tcataatctt ggatggcttt caagaaagcc tcctgatcaa aacggcggaa cataatcacg
     1021 cggagaccga ccataaagta acccaaagta atatgaaagc cgaaagcatg gaagaaaggc
     1081 aagtagacca agacggtgac accaggaatc agctgagtgc cgtagcgtgg atcgagagca
     1141 tggatcagac gcacgcaaat gttttgatgg gtctgcatga ctccctttgg gagtccagta
     1201 gtaccgctgc tacacagaat ggctgcaact tgttccacag ggtcgaagtg gagtggttta
     1261 aagtttgcga tgttgccgtc tgaatagcga gagatgaaat taggcaaaga ttcgcaaccg
     1321 tgaatattct ccacagtgtc caagatgatg atacgcttaa taaagttggt gcggctttgg
     1381 acttccagga ctttgttcag aatattctta gtggtgaaga caatctgtgg cttagagata
     1441 cccatgactt tacacagttc gtcgggaatg tagctctcgt tgactggagc cacgatcata
     1501 ccgatatacc atgcggcgat gactggaatg aagaaacggg tattgttttc agcacagata
     1561 ctaacgacgt cgttcatctt gtagccacaa ttgtggaggg actgagccag caagacggtt
     1621 gcctcaaaaa actccttgta gctcaaagat tcatcgccga ccacatcgac caaggcttga
     1681 ggcaaatgag agtgcttgcg gagagcacga aacagcattt cgccggcagt caaatcctcc
     1741 aaaggatgga gaggctcagg gccatagatg acatttttct cacgctttac catggtggct
     1801 ttaccaacag taccggaatg ccaagcttac ttagatcgca gatctcgagc ccgggctgca
     1861 ggaattcgat aggcagctgc gccacgaagc agtcacgtcc ccaagagcag cagcttggtc
     1921 cggctgactt ataaacagtg agtagcgccc gcccacttgt ttcgggcggc ccgtaaggca
     1981 cccttatctt tgaaatgcta taagatgtgt gggaaatagt aataccaggt gtacatgatt
     2041 tgtgttaagg tgtgtggtgg ttttcgaaaa acatgggtta ttggccgttt ctgtttctca
     2101 cgcccggatt atatatctgg acttgcctag gtagcatcca tggcaacgca aggccagcaa
     2161 acccactatc aagcttggca ttccggtact gttggtaaag ccaccatggt gaaacgcgaa
     2221 aagaacgtga tctacggccc agaaccactg catccactgg aagacctcac cgctggtgag
     2281 atgctcttcc gagcactgcg taaacatagt cacctccctc aagcactcgt ggacgtcgtg
     2341 ggagacgaga gcctctccta caaagaattt ttcgaagcta ctgtgctgtt ggcccaaagc
     2401 ctccataatt gtgggtacaa aatgaacgat gtggtgagca tttgtgctga gaataacact
     2461 cgcttcttta ttcctgtaat cgctgcttgg tacatcggca tgattgtcgc ccctgtgaat
     2521 gaatcttaca tcccagatga gctgtgtaag gttatgggta ttagcaaacc tcaaatcgtc
     2581 tttactacca aaaacatctt gaataaggtc ttggaagtcc agtctcgtac taacttcatc
     2641 aaacgcatca ttattctgga taccgtcgaa aacatccacg gctgtgagag cctccctaac
     2701 ttcatctctc gttacagcga tggtaatatc gctaatttca agcccttgca ttttgatcca
     2761 gtcgagcaag tggccgctat tttgtgctcc tccggcacca ctggtttgcc taaaggtgtc
     2821 atgcagactc accagaatat ctgtgtgcgt ttgatccacg ctctcgaccc tcgtgtgggt
     2881 actcaattga tccctggcgt gactgtgctg gtgtatctgc ctttctttca cgcctttggt
     2941 ttctctatta ccctgggcta tttcatggtc ggcttgcgtg tcatcatgtt tcgtcgcttc
     3001 gaccaagaag ccttcttgaa ggctattcaa gactacgagg tgcgttccgt gatcaacgtc
     3061 ccttcagtca ttttgttcct gagcaaatct cctttggttg acaagtatga tctgagcagc
     3121 ttgcgtgagc tgtgctgtgg cgctgctcct ttggccaaag aagtggccga ggtcgctgct
     3181 aagcgtctga acctccctgg tatccgctgc ggttttggtt tgactgagag cacttctgct
     3241 aacatccata gcttgcgaga cgagtttaag tctggtagcc tgggtcgcgt gactcctctt
     3301 atggctgcaa agatcgccga ccgtgagacc ggcaaagcac tgggcccaaa tcaagtcggt
     3361 gaattgtgta ttaagggccc tatggtctct aaaggctacg tgaacaatgt ggaggccact
     3421 aaagaagcca ttgatgatga tggctggctc catagcggcg acttcggtta ctatgatgag
     3481 gacgaacact tctatgtggt cgatcgctac aaagaattga ttaagtacaa aggctctcaa
     3541 gtcgcaccag ccgaactgga agaaattttg ctgaagaacc cttgtatccg cgacgtggcc
     3601 gtcgtgggta tcccagactt ggaagctggc gagttgccta gcgcctttgt ggtgaaacaa
     3661 cccggcaagg agatcactgc taaggaggtc tacgactatt tggccgagcg cgtgtctcac
     3721 accaaatatc tgcgtggcgg cgtccgcttc gtcgattcta ttccacgcaa cgttaccggt
     3781 aagatcactc gtaaagagtt gctgaagcaa ctcctcgaaa aagctggcgg ctagtaaagt
     3841 cttcatgatt atctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat
     3901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     4021 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     4081 gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg tcgacctcga gggggggccc
     4141 ggtaccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca
     4201 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4261 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4321 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4381 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4441 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4501 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4561 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4621 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4681 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4741 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4801 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4861 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4921 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4981 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5041 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5101 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5161 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5221 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5281 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5341 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5401 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5461 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5521 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5581 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5641 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5701 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5761 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5821 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5881 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5941 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6001 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6061 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6121 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6181 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6241 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6301 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     6361 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     6421 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     6481 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     6541 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     6601 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     6661 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     6721 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6781 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     6841 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6901 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6961 gtgaattgta atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgct
     7021 ctagaactag tggatccccc gatccttatc gattttacca catttgtaga ggttttactt
     7081 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     7141 gttgtt
//
