LOCUS       sequence-471621-        8381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8381)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 8381)
  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: pcDNA3-TLR2-CFP
FEATURES             Location/Qualifiers
     source          1..8381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        603..806
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     757..777
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     851..870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        851..869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3259..3261,3262..3264,3265..3978)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3304..3325)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3565..3584)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3912..3933
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3994..4012)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3995..4012)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4032..4049)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4038..4262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4308..4736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4395..4414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4605..4626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4745..4765)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4789..4999
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4811..5007
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4858..4993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4920..4939
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5074..5868
                     /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(5128..5147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5738..5757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6042..6163
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6079..6098)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6133..6152
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6212..6228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6212..6228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6225..6247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6236..6252
                     /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(6260..6266,6267..6284,6285..6290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6305..6326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6443..6460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6694..6713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7373..8164,8165..8233))
                     /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     7996..8015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8234..8338)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      841 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc
      901 atgccacata ctttgtggat ggtgtgggtc ttgggggtca tcatcagcct ctccaaggaa
      961 gaatcctcca atcaggcttc tctgtcttgt gaccgcaatg gtatctgcaa gggcagctca
     1021 ggatctttaa actccattcc ctcagggctc acagaagctg taaaaagcct tgacctgtcc
     1081 aacaacagga tcacctacat tagcaacagt gacctacaga ggtgtgtgaa cctccaggct
     1141 ctggtgctga catccaatgg aattaacaca atagaggaag attctttttc ttccctgggc
     1201 agtcttgaac atttagactt atcctataat tacttatcta atttatcgtc ttcctggttc
     1261 aagccccttt cttctttaac attcttaaac ttactgggaa atccttacaa aaccctaggg
     1321 gaaacatctc ttttttctca tctcacaaaa ttgcaaatcc tgagagtggg aaatatggac
     1381 accttcacta agattcaaag aaaagatttt gctggactta ccttccttga ggaacttgag
     1441 attgatgctt cagatctaca gagctatgag ccaaaaagtt tgaagtcaat tcagaacgta
     1501 agtcatctga tccttcatat gaagcagcat attttactgc tggagatttt tgtagatgtt
     1561 acaagttccg tggaatgttt ggaactgcga gatactgatt tggacacttt ccatttttca
     1621 gaactatcca ctggtgaaac aaattcattg attaaaaagt ttacatttag aaatgtgaaa
     1681 atcaccgatg aaagtttgtt tcaggttatg aaacttttga atcagatttc tggattgtta
     1741 gaattagagt ttgatgactg tacccttaat ggagttggta attttagagc atctgataat
     1801 gacagagtta tagatccagg taaagtggaa acgttaacaa tccggaggct gcatattcca
     1861 aggttttact tattttatga tctgagcact ttatattcac ttacagaaag agttaaaaga
     1921 atcacagtag aaaacagtaa agtttttctg gttccttgtt tactttcaca acatttaaaa
     1981 tcattagaat acttggatct cagtgaaaat ttgatggttg aagaatactt gaaaaattca
     2041 gcctgtgagg atgcctggcc ctctctacaa actttaattt taaggcaaaa tcatttggca
     2101 tcattggaaa aaaccggaga gactttgctc actctgaaaa acttgactaa cattgatatc
     2161 agtaagaata gttttcattc tatgcctgaa acttgtcagt ggccagaaaa gatgaaatat
     2221 ttgaacttat ccagcacacg aatacacagt gtaacaggct gcattcccaa gacactggaa
     2281 attttagatg ttagcaacaa caatctcaat ttattttctt tgaatttgcc gcaactcaaa
     2341 gaactttata tttccagaaa taagttgatg actctaccag atgcctccct cttacccatg
     2401 ttactagtat tgaaaatcag taggaatgca ataactacgt tttctaagga gcaacttgac
     2461 tcatttcaca cactgaagac tttggaagct ggtggcaata acttcatttg ctcctgtgaa
     2521 ttcctctcct tcactcagga gcagcaagca ctggccaaag tcttgattga ttggccagca
     2581 aattacctgt gtgactctcc atcccatgtg cgtggccagc aggttcagga tgtccgcctc
     2641 tcggtgtcgg aatgtcacag gacagcactg gtgtctggca tgtgctgtgc tctgttcctg
     2701 ctgatcctgc tcacgggggt cctgtgccac cgtttccatg gcctgtggta tatgaaaatg
     2761 atgtgggcct ggctccaggc caaaaggaag cccaggaaag ctcccagcag gaacatctgc
     2821 tatgatgcat ttgtttctta cagtgagcgg gatgcctact gggtggagaa ccttatggtc
     2881 caggagctgg agaacttcaa tccccccttc aagttgtgtc ttcataagcg ggacttcatt
     2941 cctggcaagt ggatcattga caatatcatt gactccattg aaaagagcca caaaactgtc
     3001 tttgtgcttt ctgaaaactt tgtgaagagt gagtggtgca agtatgaact ggacttctcc
     3061 catttccgtc tttttgatga gaacaatgat gctgccattc tcattcttct ggagcccatt
     3121 gagaaaaaag ccattcccca gcgcttctgc aagctgcgga agataatgaa caccaagacc
     3181 tacctggagt ggcccatgga cgaggctcag cgggaaggat tttgggtaaa tctgagagct
     3241 gcgataaagt ccctcgagat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     3301 atcctggtcg agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc
     3361 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     3421 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc
     3481 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3541 caggagcgta ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     3601 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3661 ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc
     3721 gccgacaagc agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac
     3781 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     3841 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     3901 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3961 gacgagctgt acaagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc
     4021 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4081 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4141 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4201 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4261 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4321 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4381 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4441 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4501 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4561 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4621 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4681 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4741 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4801 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4861 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4921 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4981 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     5041 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5101 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5161 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5221 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5281 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5341 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5401 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5461 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5521 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5581 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5641 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5701 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5761 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5821 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5881 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5941 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6001 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6061 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6121 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6181 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6241 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6301 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6361 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6421 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtcg acggatcggg a
//
