LOCUS       sequence-181408-        4825 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Ubiquitous expression plasmid, contains CAG promoter (CMV immediate 
            early enhancer and chicken beta actin promoter), MCS and IRES 
            controlled Histone2B-EGFP reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4825)
  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 4825)
  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: pCI H2B-GFP
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    join(19..593,594..596,597..605)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(186..203)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     413..432
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             612..989
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1002..1011
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1008..1010,1011..1013,1014..1727)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1008..1010,1011..1013,1014..1724)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1053..1074)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1314..1333)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1661..1682
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1819..1838)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1884..1939
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1938..1957)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2330..2346)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2330..2346)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2343..2365)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2354..2370
                     /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(2378..2384,2385..2402,2403..2408))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2423..2444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2503..2698
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2549..2684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2704..2838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2741..2760)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2795..2814
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2906..2923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3077..3665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3157..3176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3836..4627,4628..4696))
                     /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     4459..4478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4697..4801)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcagcccgg gggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg
       61 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
      121 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
      181 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
      241 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
      301 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
      361 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
      421 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
      481 tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
      541 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
      601 caaccatggt tatgccagag ccagcgaagt ctgctcccgc cccgaaaaag ggctccaaga
      661 aggcggtgac taaggcgcag aagaaaggcg gcaagaagcg caagcgcagc cgcaaggaga
      721 gctattccat ctatgtgtac aaggttctga agcaggtcca ccctgacacc ggcatttcgt
      781 ccaaggccat gggcatcatg aattcgtttg tgaacgacat tttcgagcgc atcgcaggtg
      841 aggcttcccg cctggcgcat tacaacaagc gctcgaccat cacctccagg gagatccaga
      901 cggccgtgcg cctgctgctg cctggggagt tggccaagca cgccgtgtcc gagggtacta
      961 aggccatcac caagtacacc agcgctaagg atccaccggt cgccaccatg gtgagcaagg
     1021 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1081 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1141 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1201 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1261 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1321 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1381 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1441 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     1501 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1561 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1621 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1681 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcaatt
     1741 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1801 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1861 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1921 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     1981 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     2041 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     2101 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     2161 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     2221 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcaccg
     2281 tcgaccagct ggtcgacggt gcaccgtcga ccagcttggc gtaatcatgg tcatagctgt
     2341 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2401 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2461 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     2521 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2581 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2641 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2701 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2761 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2821 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4801 gcacatttcc ccgaaaagtg ccacc
//
