LOCUS       sequence-208665-        5365 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5365)
  AUTHORS   Cheutin T, McNairn AJ, Jenuwein T, Gilbert DM, Singh PB, Misteli T
  TITLE     Maintenance of stable heterochromatin domains by dynamic HP1 
            binding.
  JOURNAL   Science. 2003 Jan 31. 299(5607):721-5.
  PUBMED    12560555
REFERENCE   2  (bases 1 to 5365)
  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: "Science. 
            2003 Jan 31. 299(5607):721-5."
COMMENT     Sequence Label: GFP-HP1gamma
FEATURES             Location/Qualifiers
     source          1..5365
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      918..927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             930..947
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             948..1661
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(978..1006)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1601..1621
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2196..2330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2366..2384)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2405..2421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2405..2421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2418..2440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2429..2445
                     /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(2453..2459,2460..2477,2478..2483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2498..2519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2636..2653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2807..3395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2887..2906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3566..4357,4358..4426))
                     /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     4189..4208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4427..4531)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4557..5012)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4694..4715)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4906..4925
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5139..5161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5153..5170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5154..5170
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5177..5196
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5177..5195
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5205..5339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 acgtcgcatg ctcctctagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctgg
      901 gccgctctag aactagcgcc accatgggac atcaccatca ccatcacatg agtaaaggag
      961 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     1021 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     1081 aatttatttg cactactgga aaactacctg ttccatggcc aacacttgtc actactttca
     1141 cttatggtgt tcaatgcttt tcaagatacc cagatcatat gaaacggcat gactttttca
     1201 agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa gatgacggga
     1261 actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat agaatcgagt
     1321 taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg gaatacaact
     1381 ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc aaagttaact
     1441 tcaaaattag acacaacatt gaagatggaa gcgttcaact agcagaccat tatcaacaaa
     1501 atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat
     1561 ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa
     1621 cagctgctgg gattacacat ggcatggatg aactatacaa gggatccgga aaaaaacaga
     1681 atggaaagag taaaaaagtt gaagaggcag agcctgaaga atttgtcgtg gaaaaagtac
     1741 tagatcgacg tgtagtgaat gggaaagtgg aatatttcct gaagtggaag ggatttacag
     1801 atgctgacaa tacttgggaa cctgaagaaa atttagattg tccagaattg attgaagcgt
     1861 ttcttaactc tcagaaagct ggcaaagaaa aagatggtac aaaaagaaaa tctttatctg
     1921 acagtgaatc tgatgacagc aaatcaaaga agaaaagaga tgctgctgac aaaccaagag
     1981 gatttgccag aggtcttgat cctgaaagaa taattggtgc cacagacagc agtggagaat
     2041 tgatgtttct catgaaatgg aaagattcag atgaggcaga cttggtgctg gcgaaagagg
     2101 caaatatgaa gtgtcctcaa attgtaattg ctttttatga agagagacta acttggcatt
     2161 cttgtccaga agatgaagct caataagctt aacccaactt gtttattgca gcttataatg
     2221 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2281 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gacctcgagg
     2341 gggggcccgg taccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat
     2401 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2461 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2521 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     2581 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2641 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2701 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2761 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2821 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2881 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2941 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3001 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3061 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3121 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3181 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3241 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3301 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3361 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3421 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3481 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3541 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3601 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3661 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3721 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3781 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3841 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3901 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3961 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4021 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4081 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4141 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4201 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4261 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4321 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4381 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4441 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4501 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     4561 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4621 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4681 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4741 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4801 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4861 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4921 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4981 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     5041 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5101 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5161 gacggccagt gaattgtaat acgactcact atagggcgaa ttggaacttg tttattgcag
     5221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     5341 atccccccac cgcggtggcg gcccg
//
