LOCUS       sequence-495378-        6148 bp    DNA     circular SYN 05-JAN-2026
DEFINITION  A CAGGS driven expression vector containing V5 tagged mScarlet3-S2. 
            H2B for nuclear labeling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6148)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 6148)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 5, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pCAGEN-H2B-V5-mScarlet3-S2 (JDW 1529)
FEATURES             Location/Qualifiers
     source          1..6148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1746..1770
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1789..2166
                     /codon_start=1
                     /gene="HIST1H2BB"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPSKSAPAPKKGSKKAITKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSSK"
     CDS             2182..2223
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2311..2997
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFQYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKH
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     2709..2728
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3054..3078)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3170..3189)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3235..3290
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3289..3308)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3651..3667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3651..3667)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3664..3686)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3675..3691
                     /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(3699..3705,3706..3723,3724..3729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3744..3765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3824..4019
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3870..4005
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4025..4159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4062..4081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4116..4135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4227..4244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4398..4986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4478..4497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5157..5948,5949..6017))
                     /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     5780..5799
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6018..6122)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagg
     1741 atatcacaag tttgtacaaa aaagcaggct actagtgcta gcgccaccat gcctgaaccc
     1801 tctaagtctg ctccagcccc taaaaagggt tctaagaagg ctatcactaa ggcgcagaag
     1861 aaggatggta agaagcgtaa gcgcagccgc aaggagagct attctatcta tgtgtacaag
     1921 gttctgaagc aggtccaccc cgacaccggc atctcatcca aggccatggg gatcatgaac
     1981 tccttcgtca acgacatctt cgagcgcatc gcgggcgagg cttctcgcct ggctcactac
     2041 aataagcgct cgaccatcac ctccagggag attcagacgg ctgtgcgcct gctgctgcct
     2101 ggggagctgg ctaagcatgc tgtgtccgag ggcactaagg cagttaccaa gtacactagc
     2161 tctaagggtg gtgctggtgg tggtaagcct atccctaacc ctctcctcgg tctcgattct
     2221 acgggatcca ccggtgaatt cgatggtgga ggcggttcag gcggaggtgg ctctggcggt
     2281 ggcggatcgc catcaggtac catggtgagc atggatagca ccgaggcagt gatcaaggag
     2341 ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt cgagatcgag
     2401 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgag ggtgaccaag
     2461 ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttccagta cggctccagg
     2521 gccttcacga agcaccccgc cgacatcccc gactactgga agcagtcctt ccccgagggc
     2581 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgtccgt ggcccaggac
     2641 acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac caacttccct
     2701 cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcatccac cgagcggttg
     2761 taccccgagg acgtcgtgct gaagggcgac attaagcatg ccctgcgcct gaaggacggc
     2821 ggccggtacc tggcggactt caagaccacc tacagggcca agaagcccgt gcagatgccc
     2881 ggcgccttca acatcgaccg caagttggac atcacatccc acaacgagga ctacaccgtg
     2941 gtggaacagt acgaacgctc cgtggcccgc cactccaccg gcggctccgg tggctcctaa
     3001 ctataatgag gcgcgccgtc gacgttaaca cgcgttaatg atacgtatct agaacccagc
     3061 tttcttgtac aaagtggtga tatcgcggcc gcactcctca ggtgcaggct gcctatcaga
     3121 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     3181 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     3241 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     3301 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     3361 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     3421 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     3481 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     3541 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     3601 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     3661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3781 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     3841 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3901 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3961 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4021 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4141 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4201 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4801 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5041 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5101 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5161 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5221 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5281 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5341 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5401 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5461 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5521 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5581 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5641 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5701 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5761 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5821 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5881 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5941 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6001 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6061 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6121 cgcacatttc cccgaaaagt gccacctg
//
