LOCUS       sequence-169271-        5125 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 5125)
  TITLE     TLR Signaling Plasmids
REFERENCE   2  (bases 1 to 5125)
  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: pFlag-CMV1-hMD2
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(100..119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     310..331
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     594..616
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     608..625
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     609..625
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        786..1165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1166..1369
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1320..1340
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1400..1444
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             1445..1468
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    1934..2556
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2133..2152)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2580..2600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2585..2914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2765..2900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2952..2971)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2953..2971)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2985..3001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2985..3001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2998..3020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3009..3025
                     /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(3033..3039,3040..3057,3058..3063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3078..3099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3216..3233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3387..3975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3467..3486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4146..4937,4938..5006))
                     /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     4769..4788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5007..5111)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
       61 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      181 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      301 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      361 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      421 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
      481 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
      541 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      601 acgacgttgt aaaacgacgg ccagtgccaa gctgatctat acattgaatc aatattggca
      661 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
      721 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
      781 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      841 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      901 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      961 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1021 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     1081 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     1141 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     1201 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1261 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     1321 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     1381 ccgtcagaat taattcacca tgtctgcact tctgatccta gctcttgttg gagctgcagt
     1441 tgctgactac aaagacgatg acgacaagct tgcggccgcg gaagctcaga agcagtattg
     1501 ggtctgcaac tcatccgatg caagtatttc atacacctac tgtgataaaa tgcaataccc
     1561 aatttcaatt aatgttaacc cctgtataga attgaaagga tccaaaggat tattgcacat
     1621 tttctacatt ccaaggagag atttaaagca attatatttc aatctctata taactgtcaa
     1681 caccatgaat cttccaaagc gcaaagaagt tatttgccga ggatctgatg acgattactc
     1741 tttttgcaga gctctgaagg gagagactgt gaatacaaca atatcattct ccttcaaggg
     1801 aataaaattt tctaagggaa aatacaaatg tgttgttgaa gctatttctg ggagcccaga
     1861 agaaatgctc ttttgcttgg agtttgtcat cctacaccaa cctaattcaa attagtcgac
     1921 tctagaggat cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     1981 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     2041 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     2101 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     2161 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     2221 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     2281 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     2341 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     2401 ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga cacagcatag
     2461 gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca ctgtcctctc
     2521 atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag cttggctgtg
     2581 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2641 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2701 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2761 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2821 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     2881 aggaggcttt tttggaggcc taggcttttg caaaaagctc ctcgaggaac tgaaaaacca
     2941 gaaagttaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata gctgtttcct
     3001 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3061 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3121 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3181 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5101 ggggttccgc gcacatttcc ccgaa
//
