LOCUS       sequence-192954-        5284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            mTagBFP2 in the pCS2+ expression vector. It has the SP6 RNA 
            polymerase site for in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5284)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5284)
  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            "Unpublished"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: pCS2+ H2B-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..5284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..993
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1028..1046
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1028..1045
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1097..1474
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1487..1496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2204..2230
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2234..2260
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2264..2290
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2329..2463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2353..2372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2407..2426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2576..2596)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2576..2594)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2615..2631)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2615..2631)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2628..2650)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2639..2655
                     /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(2663..2669,2670..2687,2688..2693))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2708..2729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2846..2863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3017..3605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3097..3116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3776..4567,4568..4636))
                     /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     4399..4418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4637..4741)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4767..5222)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4904..4925)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5116..5135
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttacgccagt cgaccgccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
       61 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
      121 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
      181 cggacctggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
      241 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
      301 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
      361 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
      421 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
      481 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
      541 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
      601 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
      661 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
      721 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
      781 tggcaagtac attactattg gaagtacgcc agggtacatt ggcagtactc ccattgacgt
      841 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
      901 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
      961 gaggtctata taagcaatgc tcgtttaggg aaccgccatt ctgcctgggg acgtcggagc
     1021 aagcttgatt taggtgacac tatagaatac aagctacttg ttctttttgc aggatcccat
     1081 cgacggtacc gccaccatgc cagagccagc gaagtctgct cccgccccga aaaagggctc
     1141 caagaaggcg gtgactaagg cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa
     1201 ggagagctat tccatctatg tgtacaaggt tctgaagcag gtccaccctg acaccggcat
     1261 ttcgtccaag gccatgggca tcatgaattc gtttgtgaac gacattttcg agcgcatcgc
     1321 aggtgaggct tcccgcctgg cgcattacaa caagcgctcg accatcacct ccagggagat
     1381 ccagacggcc gtgcgcctgc tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg
     1441 tactaaggcc atcaccaagt acaccagcgc taaggatcca ccggtcgcca ccatggtttc
     1501 gaagggagag gagctgatca aagagaatat gcacatgaag ctgtatatgg agggaaccgt
     1561 cgacaaccac cactttaagt gcaccagcga gggtgaggga aagccctacg agggcacaca
     1621 gaccatgcgt attaaagttg tcgagggagg tccactgcca tttgcattcg atatcctggc
     1681 caccagtttc ctctatggca gcaagacatt catcaatcat actcagggaa taccagattt
     1741 cttcaagcaa tccttccctg aaggcttcac atgggaacgc gttactacct atgaggacgg
     1801 cggagttctg actgccaccc aagatactag cctgcaggat ggttgcttga tttacaacgt
     1861 gaagatacgc ggcgtgaact ttacctcgaa cggcccggtt atgcagaaaa agacgctcgg
     1921 atgggaggcc tttacggaga cgctgtaccc cgccgatggc ggtcttgaag gtcgcaacga
     1981 tatggctctg aagctggtgg gcggttcgca ccttatagcc aatgccaaga ctacgtaccg
     2041 tagtaagaag ccagcgaaga acctgaagat gcccggtgtg tactacgtgg actataggct
     2101 ggagcgcatt aaggaagcta acaacgaaac ctacgtcgag cagcacgaag tcgccgtcgc
     2161 ccgctactgt gatttgccta gtaagttggg tcataagctc aactacccat acgatgttcc
     2221 tgactatgcg ggctatccct atgacgtccc ggactatgca ggatatccat atgacgttcc
     2281 agattacgct ggcggataat ctagaactat agtgagtcgt attacgtaga tccagacatg
     2341 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2401 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2461 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2521 ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt
     2581 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     2641 gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg
     2701 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     2761 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2821 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3001 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3061 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3121 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3181 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3241 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3301 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3361 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3421 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3481 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3541 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3601 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3661 gttaagggat tttggtcatg agattatcaa aaatgatctt cacctagatc cttttaaatt
     3721 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3781 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3841 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3901 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3961 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     4021 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4081 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4141 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4201 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     4261 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4321 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4381 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4441 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4501 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4561 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4621 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4681 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4741 gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc
     4801 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4861 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4921 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4981 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     5041 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     5101 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5161 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5221 gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5281 gcta
//
