LOCUS       sequence-487265-        6652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of H2B-mTagBFP2 for nuclei isolation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6652)
  TITLE     Maria J Galazo Lab Materials
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6652)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAV-CAG-H2B-mTagBFP2-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..6652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        161..540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        542..819
                     /label=chicken beta-actin promoter
     intron          820..1836
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1760..1782
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1844..1863
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1890..1914
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1921..2298
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     regulatory      2311..2320
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2317..3027
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    complement(3031..3055)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3065..3653
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3118..3138)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3536..3547)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3683..3700)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3689..3896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3904..4044
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3904..4033
                     /label=AAV2 ITR
     rep_origin      4119..4574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4206..4225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4416..4437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4591..4610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4710..4732
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4770..4788)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4856..4960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4961..5029,5030..5821)
                     /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     complement(5179..5198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5992..6580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6481..6500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6642..6652,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgctc gacattgatt attgactagt
      181 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      241 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      301 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      361 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      421 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      481 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      541 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
      601 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
      661 gggcgcgcgc caggcggggc ggggcggggc grggggcggg gcggggcgag gcggagaggt
      721 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      781 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      841 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      901 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      961 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
     1021 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
     1081 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
     1141 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1201 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     1261 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     1321 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     1381 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1441 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1501 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1561 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1621 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1681 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1741 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1801 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1861 gtgctgtctc atcattttgg caaagaattg caagtttgta caaaaaagca ggctgccacc
     1921 atgccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
     1981 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
     2041 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
     2101 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc
     2161 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
     2221 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccgtcacc
     2281 aagtacacca gcgctaagga cccaccggtc gccaccatgg tgtctaaggg cgaagagctg
     2341 attaaggaga acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc
     2401 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     2461 gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac
     2521 ggcagcaaga ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc
     2581 cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     2641 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     2701 aacttcacat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc
     2761 gagacgctgt accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc
     2821 gtgggcggga gccatctgat cgcaaacgcc aagaccacat atagatccaa gaaacccgct
     2881 aagaacctca agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag
     2941 gccaacaacg agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc
     3001 cctagcaaac tggggcacaa gcttaattaa acccagcttt cttgtacaaa gtgggaattc
     3061 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3121 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3181 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3241 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3301 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3361 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3421 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3481 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3541 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3601 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcggga
     3661 attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3721 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3781 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3841 gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagggc
     3901 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3961 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4021 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4081 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4141 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     4201 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4261 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4321 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4381 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4441 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     4501 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4561 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4621 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4681 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4741 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4801 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5101 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5161 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5221 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5281 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5341 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5401 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5461 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5521 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5581 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5641 gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact
     5701 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5761 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5821 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5881 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5941 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6001 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6061 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6121 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6181 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6241 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6301 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6361 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6421 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6481 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6541 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6601 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc ag
//
