LOCUS       sequence-7721-ex        8567 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 8567)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 8567)
  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-TLR10-YFP
FEATURES             Location/Qualifiers
     source          1..8567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3384..3386,3387..3389,3390..4103)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3429..3450)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3690..3709)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4037..4058
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4119..4137)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4120..4137)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4157..4174)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4163..4387
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4433..4861
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4520..4539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4730..4751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4870..4890)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4875..5205
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5056..5191
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5118..5137
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5272..6066
                     /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(5326..5345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5936..5955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6240..6361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6277..6296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6331..6350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6410..6426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6410..6426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6423..6445)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6434..6450
                     /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(6458..6464,6465..6482,6483..6488))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6503..6524
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6641..6658)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6812..7400)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6892..6911)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7571..8362,8363..8431))
                     /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     8194..8213
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8432..8536)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtcc agtgtggtgg aattgccctt caccatgaga ctcatcagaa
      961 acatttacat attttgtagt attgttatga cagcagaggg tgatgctcca gagctgccag
     1021 aagaaaggga actgatgacc aactgctcca acatgtctct aagaaaggtt cccgcagact
     1081 tgaccccagc cacaacgaca ctggatttat cctataacct cctttttcaa ctccagagtt
     1141 cagattttca ttctgtctcc aaactgagag ttttgattct atgccataac agaattcaac
     1201 agctggatct caaaaccttt gaattcaaca aggagttaag atatttagat ttgtctaata
     1261 acagactgaa gagtgtaact tggtatttac tggcaggtct caggtattta gatctttctt
     1321 ttaatgactt tgacaccatg cctatctgtg aggaagctgg caacatgtca cacctggaaa
     1381 tcctaggttt gagtggggca aaaatacaaa aatcagattt ccagaaaatt gctcatctgc
     1441 atctaaatac tgtcttctta ggattcagaa ctcttcctca ttatgaagaa ggtagcctgc
     1501 ccatcttaaa cacaacaaaa ctgcacattg ttttaccaat ggacacaaat ttctgggttc
     1561 ttttgcgtga tggaatcaag acttcaaaaa tattagaaat gacaaatata gatggcaaaa
     1621 gccaatttgt aagttatgaa atgcaacgaa atcttagttt agaaaatgct aagacatcgg
     1681 ttctattgct taataaagtt gatttactct gggacgacct tttccttatc ttacaatttg
     1741 tttggcatac atcagtggaa cactttcaga tccgaaatgt gacttttggt ggtaaggctt
     1801 atcttgacca caattcattt gactactcaa atactgtaat gagaactata aaattggagc
     1861 atgtacattt cagagtgttt tacattcaac aggataaaat ctatttgctt ttgaccaaaa
     1921 tggacataga aaacctgaca atatcaaatg cacaaatgcc acacatgctt ttcccgaatt
     1981 atcctacgaa attccaatat ttaaattttg ccaataatat cttaacagac gagttgttta
     2041 aaagaactat ccaactgcct cacttgaaaa ctctcatttt gaatggcaat aaactggaga
     2101 cactttcttt agtaagttgc tttgctaaca acacaccctt ggaacacttg gatctgagtc
     2161 aaaatctatt acaacataaa aatgatgaaa attgctcatg gccagaaact gtggtcaata
     2221 tgaatctgtc atacaataaa ttgtctgatt ctgtcttcag gtgcttgccc aaaagtattc
     2281 aaatacttga cctaaataat aaccaaatcc aaactgtacc taaagagact attcatctga
     2341 tggccttacg agaactaaat attgcattta attttctaac tgatctccct ggatgcagtc
     2401 atttcagtag actttcagtt ctgaacattg aaatgaactt cattctcagc ccatctctgg
     2461 attttgttca gagctgccag gaagttaaaa ctctaaatgc gggaagaaat ccattccggt
     2521 gtacctgtga attaaaaaat ttcattcagc ttgaaacata ttcagaggtc atgatggttg
     2581 gatggtcaga ttcatacacc tgtgaatacc ctttaaacct aaggggaact aggttaaaag
     2641 acgttcatct ccacgaatta tcttgcaaca cagctctgtt gattgtcacc attgtggtta
     2701 ttatgctagt tctggggttg gctgtggcct tctgctgtct ccactttgat ctgccctggt
     2761 atctcaggat gctaggtcaa tgcacacaaa catggcacag ggttaggaaa acaacccaag
     2821 aacaactcaa gagaaatgtc cgattccacg catttatttc atacagtgaa catgattctc
     2881 tgtgggtgaa gaatgaattg atccccaatc tagagaagga agatggttct atcttgattt
     2941 gcctttatga aagctacttt gaccctggca aaagcattag tgaaaatatt gtaagcttca
     3001 ttgagaaaag ctataagtcc atctttgttt tgtctcccaa ctttgtccag aatgagtggt
     3061 gccattatga attctacttt gcccaccaca atctcttcca tgaaaattct gatcatataa
     3121 ttcttatctt actggaaccc attccattct attgcattcc caccaggtat cataaactga
     3181 aagctctcct ggaaaaaaaa gcatacttgg aatggcccaa ggataggcgt aaatgtgggc
     3241 ttttctgggc aaaccttcga gctgctatta atgttaatgt attagccacc agagaaatgt
     3301 atgaactgca gacattcaca gagttaaatg aagagtctcg aggttctaca atctctctga
     3361 tgagaacaga ttgtctagtc gagatggtga gcaagggcga ggagctgttc accggggtgg
     3421 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     3481 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     3541 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg aagtgcttcg
     3601 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     3661 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     3721 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3781 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     3841 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     3901 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3961 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     4021 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4081 gcatggacga gctgtacaag taatctagag ggccctattc tatagtgtca cctaaatgct
     4141 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4201 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4261 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4321 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4381 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4441 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4501 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4561 ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta
     4621 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4681 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4741 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4801 ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4861 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc
     4921 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5041 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5101 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5161 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5221 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5281 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5341 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5401 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5461 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5521 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5581 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5641 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5701 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5761 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5821 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5881 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5941 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6001 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6061 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6121 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6181 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6361 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6421 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6481 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6541 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     7021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     7261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7321 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7381 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7441 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7501 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7561 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7621 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7681 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7741 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7801 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7861 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7921 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7981 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8041 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8101 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8161 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8221 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8281 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8341 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8401 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8461 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8521 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
