LOCUS       sequence-192965-        5276 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Contains a fusion of human histone H2B to fluorescent protein 
            mCerulean 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 5276)
  TITLE     Dr. Philipp Keller Lab
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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            "Unpublished"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-mCerulean
FEATURES             Location/Qualifiers
     source          1..5276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..1001
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1036..1054
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1036..1053
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1105..1482
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      1495..1504
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1501..1503,1504..1506,1507..2217)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1546..1567)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1807..1826)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2154..2175
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2221..2286
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     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 cttcgctatt acgccagtcg accgccaatt caatatggcg tatatggact catgccaatt
       61 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
      121 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
      181 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
      241 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
      301 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
      361 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
      421 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
      481 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
      541 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
      601 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
      661 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
      721 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
      781 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
      841 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
      901 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
      961 cctaatggga ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac
     1021 gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag
     1081 gatcccatcg acggtaccgc caccatgcca gagccagcga agtctgctcc cgccccgaaa
     1141 aagggctcca agaaggcggt gactaaggcg cagaagaaag gcggcaagaa gcgcaagcgc
     1201 agccgcaagg agagctattc catctatgtg tacaaggttc tgaagcaggt ccaccctgac
     1261 accggcattt cgtccaaggc catgggcatc atgaattcgt ttgtgaacga cattttcgag
     1321 cgcatcgcag gtgaggcttc ccgcctggcg cattacaaca agcgctcgac catcacctcc
     1381 agggagatcc agacggccgt gcgcctgctg ctgcctgggg agttggccaa gcacgccgtg
     1441 tccgagggta ctaaggccat caccaagtac accagcgcta aggatccacc ggtcgccacc
     1501 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1561 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1621 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1681 ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     1741 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1801 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1861 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1921 aagctggagt acaacgccat cagcgacaac gtctatatca ccgccgacaa gcagaagaac
     1981 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2041 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2101 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2161 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagatg
     2221 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     2281 gacaagtaac gtaagctagt 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 aaaggatctt 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 gggcct
//
