LOCUS       sequence-8702-ex        4292 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 4292)
  TITLE     Cecilia Lo plasmids
REFERENCE   2  (bases 1 to 4292)
  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: pCMV3XFLAGm43CT
FEATURES             Location/Qualifiers
     source          1..4292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             839..904
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1431..1565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1455..1474)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1509..1528
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1676..1692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1676..1692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1689..1711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1700..1716
                     /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(1724..1730,1731..1748,1749..1754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1769..1790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1907..1924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2078..2666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2158..2177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2837..3628,3629..3697))
                     /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     3460..3479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3698..3802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3870..3888
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3926..3948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4048..4067
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4261..4283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4275..4292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4276..4292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tcgaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg gccgcaattc ccgggatgga
      841 ctacaaagac catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga
      901 caagcttgtt aaggatcgcg tgaagggaag aagcgatcct taccacgcca ccaccggccc
      961 actgagccca tccaaagact gcggatctcc aaaatatgct tacttcaatg gctgctcctc
     1021 accaacggcc ccactctcac ctatgtctcc tcctgggtac aagctggtca ctggtgacag
     1081 aaacaattcc tcctgccgca attacaacaa gcaagccagc gagcaaaact gggcgaatta
     1141 cagcgcagag caaaatcgaa tggggcaggc cggaagcacc atctccaact cccacgccca
     1201 gccgtttgat ttccctgacg acagccaaaa tgccaaaaaa gttgctgctg gacacgaact
     1261 ccagccctta gctatcgtgg atcagcgacc ttccagcaga gccagcagcc gcgccagcag
     1321 cagacctcgg cctgatgacc tggagattta aacaggcttg gaattcatcg atagatctga
     1381 ttttggtctg ctttctgaca aactcggcct cgactctagg cggccgcggg gatccagaca
     1441 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1501 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1561 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1621 ttttttcgga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
     1681 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1741 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1801 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1861 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     2521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3121 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3901 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3961 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4021 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4081 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     4141 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4201 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4261 cccagtcacg acgttgtaaa acgacggcca gt
//
