LOCUS       sequence-181397-        3755 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CAG-driven ubiquitous expression of MCP-VP64. MCP is bacteriophage 
            MS2 coat protein, VP64 is transcriptional activation domain. Part of
            SAM-mediated gene activation system in chicken embryos..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3755)
  AUTHORS   Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, 
            Sauka-Spengler T
  TITLE     Genome and epigenome engineering CRISPR toolkit for in vivo 
            modulation of cis-regulatory interactions and gene expression in the
            chicken embryo.
  JOURNAL   Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 
            10.1242/dev.160333.
  PUBMED    29386245
REFERENCE   2  (bases 1 to 3755)
  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"; journalName: 
            "Development."; date: "2018-02-23"; volume: "145(4). pii: 
            dev.160333. doi"; pages: " 10.1242/dev.160333"
COMMENT     Sequence Label: pCAG MCP-VP64
FEATURES             Location/Qualifiers
     source          1..3755
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             8..394
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             449..469
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             485..634
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(743..762)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    808..863
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(862..881)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1254..1270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1254..1270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1267..1289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1278..1294
                     /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(1302..1308,1309..1326,1327..1332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1347..1368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1427..1622
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1473..1608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1628..1762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1665..1684)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1719..1738
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1830..1847)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2001..2589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2081..2100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2760..3551,3552..3620))
                     /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     3383..3402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3621..3725)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caccatggct tcaaacttta ctcagttcgt gctcgtggac aatggtggga caggggatgt
       61 gacagtggct ccttctaatt tcgctaatgg ggtggcagag tggatcagct ccaactcacg
      121 gagccaggcc tacaaggtga catgcagcgt caggcagtct agtgcccaga agagaaagta
      181 taccatcaag gtggaggtcc ccaaagtggc tacccagaca gtgggcggag tcgaactgcc
      241 tgtcgccgct tggaggtcct acctgaacat ggagctcact atcccaattt tcgctaccaa
      301 ttctgactgt gaactcatcg tgaaggcaat gcaggggctc ctcaaagacg gtaatcctat
      361 cccttccgcc atcgccgcta actcaggtat ctacagcgct ggaggaggtg gaagcggagg
      421 aggaggaagc ggaggaggag gtagcggacc taagaaaaag aggaaggtgg gttccggacg
      481 ggctgacgca ttggacgatt ttgatctgga tatgctggga agtgacgccc tcgatgattt
      541 tgaccttgac atgcttggtt cggatgccct tgatgacttt gacctcgaca tgctcggcag
      601 tgacgccctt gatgatttcg acctggacat gctgggccgc aattcactcc tcaactgata
      661 atgaactttg tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
      721 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
      781 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
      841 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
      901 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
      961 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     1021 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     1081 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     1141 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     1201 accgtcgacc agctggtcga cggtgcaccg tcgaccagct tggcgtaatc atggtcatag
     1261 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1321 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     1441 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     1501 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     1561 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1621 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1681 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1741 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     1801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3181 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3241 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3301 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3361 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3421 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3481 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3541 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3601 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3661 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3721 ccgcgcacat ttccccgaaa agtgccacct gggtc
//
