LOCUS       sequence-192973-        5230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            TagRFP-T 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 5230)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5230)
  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            "Unpublished"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-TagRFP-T
FEATURES             Location/Qualifiers
     source          1..5230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..991
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1026..1044
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1026..1043
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1095..1472
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1485..1494
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2273..2407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2297..2316)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2351..2370
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2520..2540)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2520..2538)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2559..2575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2559..2575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2572..2594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2583..2599
                     /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(2607..2613,2614..2631,2632..2637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2652..2673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2790..2807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2961..3549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3041..3060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3720..4511,4512..4580))
                     /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     4343..4362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4581..4685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4711..5166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4848..4869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5060..5079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt caatatggtg
       61 gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg
      121 gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg
      181 gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg
      241 ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc
      301 atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata
      361 tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat
      421 gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg
      481 ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt
      541 ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac
      601 gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca
      661 ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg
      721 acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg
      781 gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc attgacgtca
      841 atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg
      901 ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga
      961 ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac gtcggagcaa
     1021 gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag gatcccatcg
     1081 acggtaccgc caccatgcca gagccagcga agtctgctcc cgccccgaaa aagggctcca
     1141 agaaggcggt gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc agccgcaagg
     1201 agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac accggcattt
     1261 cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag cgcatcgcag
     1321 gtgaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc agggagatcc
     1381 agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg tccgagggta
     1441 ctaaggccat caccaagtac accagcgcta aggatccacc ggtcgccacc atggtgtcaa
     1501 aaggtgaaga actgattaag gaaaatatgc acatgaagtt atacatggaa ggcacggtca
     1561 ataatcacca ctttaaatgt acatctgaag gagagggaaa accgtacgaa ggaactcaga
     1621 ctatgcgaat caaagtcgtg gagggtggcc cgctgccttt cgccttcgat attttggcta
     1681 ctagcttcat gtacggatct cgcaccttta ttaaccatac ccagggaatc ccagatttct
     1741 ttaaacaatc gttcccggaa ggtttcacgt gggaaagagt gacgacatac gaagatggag
     1801 gagtcctcac cgctacccag gacacgagtt tgcaagatgg atgcttgatc tatgatgtaa
     1861 agattcgagg cgtaaatttc ccttcgaacg gcccagtcat gcaaaagaag actttgggat
     1921 gggaagcaaa tactgaaatg ctttatcctg ctgatggagg tcttgaagga cgtacggata
     1981 tggcactgaa actggtgggc ggaggacact tgatatgtaa tttcaaaact acgtatcgaa
     2041 gtaaaaagcc ggccaagaat ctgaaaatgc caggagttta ttacgttgac cacagacttg
     2101 aaagaattaa ggaggcagat aaagaaacgt acgtggaaca acacgaagta gccgttgctc
     2161 gctattgtga cttgccatcg aaacttggac ataaacttaa tggaatggat gaactttata
     2221 aatagagcgg cccctctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga
     2281 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2341 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2401 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2461 ggttttttaa ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc
     2521 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     2581 aattgttatc cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc
     2641 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2701 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4681 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4741 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4801 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4861 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4921 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4981 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5041 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5101 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5161 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5221 cttcgctatt
//
