LOCUS       sequence-173543-        6881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding Histone 2B GFP fusion and 
            miniSOG2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6881)
  AUTHORS   Makhijani K, To TL, Ruiz-Gonzalez R, Lafaye C, Royant A, Shu X
  TITLE     Precision Optogenetic Tool for Selective Single- and Multiple-Cell 
            Ablation in a Live Animal Model System.
  JOURNAL   Cell Chem Biol. 2017 Jan 19;24(1):110-119. doi: 
            10.1016/j.chembiol.2016.12.010. Epub 2017 Jan 5.
  PUBMED    28065655
REFERENCE   2  (bases 1 to 6881)
  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"; doi: 
            "10.1016/j.chembiol.2016.12.010"; journalName: "Cell Chem Biol"; 
            date: "2017-01-19- 19"; volume: "24"; issue: "1"; pages: "110-119"
COMMENT     Sequence Label: pcDNA3.1_miniSOG2 T2A H2B-EGFP
FEATURES             Location/Qualifiers
     source          1..6881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      923..932
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1253..1306
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1313..1690
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             join(1724..1726,1727..1729,1730..2440)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1769..1790)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2030..2049)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2377..2398
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2472..2489)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2478..2702
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2748..3176
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2835..2854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3045..3066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3185..3205)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3190..3519
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3370..3505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3432..3451
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3586..4380
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3640..3659)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4250..4269
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4554..4675
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4591..4610)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4645..4664
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4724..4740)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4724..4740)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4737..4759)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4748..4764
                     /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(4772..4778,4779..4796,4797..4802))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4817..4838
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4955..4972)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5126..5714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5206..5225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5885..6676,6677..6745))
                     /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     6508..6527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6746..6850)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgccaccat ggaaaaaagc ttcgtgatta ctgacccgcg
      961 cctgccggat aatccgatca tcttcgcctc cgatagtttc ctggaactga ctgaatacag
     1021 ccgtgaggaa attctgggtc gtaatccccg tttcctgcgg ggcccggaaa ccgatcaagc
     1081 aactgttcag aaaattcatg atgcaatccg tgatcaacgt gaaatcaccg tgcagctgat
     1141 taactatacc aaaagcggca agaaattctg gaacttgttt cgcctgcagc cgattcgcga
     1201 tcagaaaggt gagctgcagt acttcatcgg ggttcagctg gatggtggat ccgagggcag
     1261 aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccagaat tgatgccaga
     1321 gccagcgaag tctgctcccg ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca
     1381 gaagaaaggc ggcaagaagc gcaagcgcag ccgcaaggag agctattcca tctatgtgta
     1441 caaggttctg aagcaggtcc accctgacac cggcatttcg tccaaggcca tgggcatcat
     1501 gaattcgttt gtgaacgaca ttttcgagcg catcgcaggt gaggcttccc gcctggcgca
     1561 ttacaacaag cgctcgacca tcacctccag ggagatccag acggccgtgc gcctgctgct
     1621 gcctggggag ttggccaagc acgccgtgtc cgagggtact aaggccatca ccaagtacac
     1681 cagcgctaag ctcgagtctg gtggcgggag tggtggcggg tccatggtga gcaagggcga
     1741 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1801 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1861 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1921 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1981 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2041 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2101 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2161 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggccaactt
     2221 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2281 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2341 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2401 cgccgccggg atcactctcg gcatggacga gctgtacaag tctagagggc ccgtttaaac
     2461 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2521 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2581 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2641 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2701 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2821 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2941 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3061 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3181 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3241 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3361 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3421 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3481 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3541 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3601 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3661 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3721 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3781 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3841 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3901 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3961 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4021 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4081 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4141 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4201 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4261 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4321 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4381 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4441 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4501 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4561 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4621 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4681 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4741 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4801 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4861 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4921 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4981 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5041 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5101 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5161 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5221 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5281 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5341 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5401 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5461 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5521 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5581 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5641 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5701 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5761 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5821 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5881 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5941 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6001 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6061 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6121 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6181 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6241 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6301 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6361 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6421 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6481 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6541 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6601 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6661 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6721 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6781 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6841 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
