LOCUS       sequence-7425-ex        6807 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 6807)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 6807)
  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-TRAM-CFP
FEATURES             Location/Qualifiers
     source          1..6807
                     /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(1624..1626,1627..1629,1630..2343)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1669..1690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1930..1949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2277..2298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2359..2377)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2360..2377)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2397..2414)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2403..2627
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2673..3101
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2760..2779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2970..2991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3110..3130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3115..3445
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3296..3431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3358..3377
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3512..4306
                     /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(3566..3585)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4176..4195
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4480..4601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4517..4536)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4571..4590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4650..4666)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4650..4666)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4663..4685)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4674..4690
                     /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(4698..4704,4705..4722,4723..4728))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4743..4764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4881..4898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5052..5640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5132..5151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5811..6602,6603..6671))
                     /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     6434..6453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6672..6776)
                     /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 ccatgggtat cgggaagtct aaaataaatt cctgccctct ttctctctct
      961 tggggtaaaa ggcacagtgt ggatacaagt ccaggatatc atgagtcaga ttccaagaag
     1021 tctgaagatc tatccttgtg taatgttgct gagcacagca atacaacaga ggggccaaca
     1081 ggaaagcagg agggagctca gagcgtggaa gagatgtttg aagaagaagc tgaagaagag
     1141 gtgttcctca aatttgtgat attgcatgca gaagatgaca cagatgaagc cctcagagtc
     1201 cagaatctgc tacaagatga ctttggtatc aaacccggaa taatctttgc tgagatgcca
     1261 tgtggcagac agcatttaca gaatttagat gatgctgtaa atgggtctgc atggacaatc
     1321 ttattactga ctgaaaactt tttaagagat acttggtgta atttccagtt ctatacgtcc
     1381 ctaatgaact ccgttaacag gcagcataaa tacaactctg ttatacccat gcggcccctg
     1441 aacaatcccc ttccccgaga aaggactccc tttgccctcc aaaccatcaa tgccttagag
     1501 gaagaaagtc gtggatttcc tacacaagta gaaagaattt ttcaggagtc tgtgtataag
     1561 acacaacaaa ctatatggaa agagacaaga aatatggtac aaagacaatt tattgccctc
     1621 gagatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1681 gacggcgacg taaacggcca caggttcagc gtgtccggcg agggcgaggg cgatgccacc
     1741 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1801 accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg
     1861 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgtaccatc
     1921 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1981 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2041 cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag
     2101 aacggcatca aggcccactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     2161 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2221 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2281 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2341 taatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     2401 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2461 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2521 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2581 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2641 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2821 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2881 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3001 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     3061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3121 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
     3181 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3241 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3301 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3361 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3421 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3481 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3541 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3601 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3661 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3721 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3781 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3841 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3901 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3961 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4021 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4081 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4141 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4201 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4261 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4321 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4381 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4441 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4501 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4561 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4621 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4681 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4741 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4801 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4861 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5221 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5461 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6661 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6781 tttccccgaa aagtgccacc tgacgtc
//
