LOCUS       sequence-257700-        5286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To make synthetic mRNA encoding a nuclear EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5286)
  TITLE     CS2-H2B-EGFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5286)
  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            "Unpublished"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: CS2-H2B-EGFP
FEATURES             Location/Qualifiers
     source          1..5286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             80..457
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             join(470..472,473..475,476..1189)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(470..472,473..475,476..1186)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(515..536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(776..795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1123..1144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1191..1215)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1260..1394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1284..1303)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1338..1357
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1507..1527)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1507..1525)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1546..1562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1546..1562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1559..1581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1570..1586
                     /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(1594..1600,1601..1618,1619..1624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1639..1660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1777..1794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1948..2536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2028..2047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2707..3498,3499..3567))
                     /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     3330..3349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3568..3672)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3698..4153)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3835..3856)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4047..4066
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4224..5208
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        5243..5261
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5243..5260
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ggatcccatc gattcgaatt caaggcctct cgagcctcta gaactatagt gagtcgtatt
       61 actcgacggt accgccacca tgccagagcc agcgaagtct gctcccgccc cgaaaaaggg
      121 ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc aagaagcgca agcgcagccg
      181 caaggagagc tattccatct atgtgtacaa ggttctgaag caggtccacc ctgacaccgg
      241 catttcgtcc aaggccatgg gcatcatgaa ttcgtttgtg aacgacattt tcgagcgcat
      301 cgcaggtgag gcttcccgcc tggcgcatta caacaagcgc tcgaccatca cctccaggga
      361 gatccagacg gccgtgcgcc tgctgctgcc tggggagttg gccaagcacg ccgtgtccga
      421 gggtactaag gccatcacca agtacaccag cgctaaggat ccaccggtca tggtgagcaa
      481 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
      541 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
      601 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
      661 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
      721 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
      781 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
      841 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
      901 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
      961 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     1021 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1081 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1141 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat acccagcttt
     1201 cttgtacaaa gtggttgatc ctctcgagcc tctagaacta tagtgagtcg tattacgtag
     1261 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     1321 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     1381 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     1441 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
     1501 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     1561 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
     1621 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     1681 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1741 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     1801 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1861 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2761 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     2821 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     2881 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     2941 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3001 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3061 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3121 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3181 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3241 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3301 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3361 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3421 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3481 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3541 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3601 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3661 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     3721 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3781 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3841 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3901 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3961 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4021 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4081 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4141 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     4201 cgggcctctt cgctattacg ccagtcgacc gccaattcaa tatggcgtat atggactcat
     4261 gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat atggactcgt
     4321 gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac ttggcaccat
     4381 gccaattcaa tatggcggac ctggcactgt gccaactggg gaggggtcta cttggcacgg
     4441 tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga attggcatgg
     4501 tgccaataat ggcggccata ttggctatat gccaggatca atatataggc aatatccaat
     4561 atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg ccctatgcca
     4621 tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca atgggcggtc
     4681 ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca ttgacgtcaa
     4741 tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt gacgtatatg
     4801 gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat gcccattgac
     4861 gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat tcatatccgt
     4921 tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt acatcaatat
     4981 ctattaatag taacttggca agtacattac tattggaagt acgccagggt acattggcag
     5041 tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt acatccccat
     5101 tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta aatgggcggt
     5161 aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg ccattctgcc
     5221 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt
     5281 tttgca
//
