LOCUS       sequence-192968-        5215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            mEmerald 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 5215)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5215)
  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            "Unpublished"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-mEmerald
FEATURES             Location/Qualifiers
     source          1..5215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..992
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1027..1045
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1027..1044
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1096..1473
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1486..1495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1492..1494,1495..1497,1498..2211)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1537..1558)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1798..1817)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2145..2166
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2259..2393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2283..2302)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2337..2356
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2506..2526)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2506..2524)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2545..2561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2545..2561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2558..2580)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2569..2585
                     /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(2593..2599,2600..2617,2618..2623))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2638..2659
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2776..2793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2947..3535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3027..3046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3706..4497,4498..4566))
                     /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     4329..4348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4567..4671)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4697..5152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4834..4855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5046..5065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tacgccagtc gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
       61 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
      121 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
      181 ggacctggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
      241 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
      301 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
      361 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
      421 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
      481 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
      541 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
      601 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
      661 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
      721 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
      781 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
      841 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
      901 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
      961 aggtctatat aagcaatgct cgtttaggga accgccattc tgcctgggga cgtcggagca
     1021 agcttgattt aggtgacact atagaataca agctacttgt tctttttgca ggatcccatc
     1081 gacggtaccg ccaccatgcc agagccagcg aagtctgctc ccgccccgaa aaagggctcc
     1141 aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg cagccgcaag
     1201 gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga caccggcatt
     1261 tcgtccaagg ccatgggcat catgaattcg tttgtgaacg acattttcga gcgcatcgca
     1321 ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc cagggagatc
     1381 cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt gtccgagggt
     1441 actaaggcca tcaccaagta caccagcgct aaggatccac cggtcgccac catggtgagc
     1501 aagggcgagg agctattcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1561 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1621 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1681 accttgacct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     1741 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1801 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1861 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1921 tacaactaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
     1981 gtgaacttca agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2041 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2101 acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2161 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggcccc
     2221 tctcgagcct ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     2281 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2341 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2401 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     2461 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     2521 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2581 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     2641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3361 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3661 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3841 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4561 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4681 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     4741 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4801 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4861 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4921 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4981 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5041 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5101 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     5161 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctat
//
